Sodium channel drugs and uses

ABSTRACT

The compounds of this invention comprise 2-10 ligands covalently connected, each of the ligands being capable of binding to a ligand binding site in a Na +  channel, thereby modulating the biological activities thereof.

This is a continuation-in-part application of application U.S. Ser. No.09/325,563, filed Jun. 4, 1999 and now abandoned, which claims thebenefit of U.S. Provisional Application No. 60/088,465 filed Jun. 8,1998 and No. 60/093,068 filed Jul. 16, 1998, the entire contents ofwhich are incorporated herein by reference.

BACKGROUND

1. Field of the Invention

This invention relates to novel multibinding compounds that bind tosodium (Na⁺) channels and modulate their activity. The compounds of thisinvention comprise 2-10 Na⁺ channel ligands covalently connected by alinker or linkers, wherein the ligands in their monovalent (i.e.,unlinked) state bind to and are capable of modulating the activity ofone or more types of Na⁺ channel. The manner of linking the ligandstogether is such that the multibinding agents thus formed demonstrate anincreased biologic and/or therapeutic effect as compared to the samenumber of unlinked ligands made available for binding to the Na⁺channel. The invention also relates to methods of using such compoundsand to methods of preparing them.

The compounds of this invention are particularly useful for treatingdiseases and conditions of mammals that are mediated by Na⁺ channels.Accordingly, this invention also relates to pharmaceutical compositionscomprising a pharmaceutically acceptable excipient and an effectiveamount of a compound of this invention.

2. State of the Art

Voltage-gated ion channels play a critical role in shaping theelectrical activity of neuronal and muscle cells, and in controlling thesecretion of neurotransmitters and hormones through the gating ofcalcium ion entry. Large families of voltage-gated sodium (Na⁺),potassium (K⁺) and calcium (Ca²⁺) ion channels have been defined usingelectrophysiological, pharmacological and molecular techniques; they arenamed according to their selective permeability for a particular cationwith reference to their voltage dependence, kinetic behavior ormolecular identity.

Although the structures of Na⁺, K⁺ and Ca²⁺ channels are quitedifferent, there are common functional elements represented in each. Thechannels are all transmembrane proteins with an ion-selective aqueouspore that, when open, extends across the membrane. Channel opening andclosing (gating) is controlled by a voltage-sensitive region of theprotein containing charged amino acids that move within the electricfield. The movement of these charged groups leads to conformationalchanges in the structure of the channel resulting in conducting(open/activated) or nonconducting (closed/inactivated) states.

Voltage-gated Na⁺ channels mediate regenerative inward currents that areresponsible for the initial depolarization of action potentials in brainneurons. Na⁺ channels are large glycoproteins that consist of varioussubunits, the principal one being the alpha (α) subunit. Na⁺ channelsexist as dimers in cardiac and skeletal muscles and exist asheterotrimers in neuronal cells. FIG. 1A shows that the α subunit has amodular architecture; it consists of four internally homologous domains(labeled I-IV), each of which contains six transmembrane segments.Prominant phosphorylation sites of the α subunit are also shown. Thefour domains fold together so as to create a central pore whosestructural constituents determine the selectivity and conductanceproperties of the channel as shown in FIG. 1B. Auxiliary beta (β)subunits are important modulators of Na⁺ channel function. Biochemicalstudies reveal the existence of two distinct β subunits (β1 and β2)associated with the brain Na⁺ channel. It should be understood that, forpurposes of simplification, other subunits that may be involved in orrequired for transporter activity have been omitted from the diagrams.

Na⁺ channels can exist in multiple ion conducting (open) andnonconducting (closed/inactivated) conformations. FIG. 2A illustrateshow Na⁺ channels open and then rapidly inactivate following voltagestimulation. Transitions between these states occurs in a voltage andtime-dependent manner. The time course and voltage dependency ofNa⁺-channel activity can be described by separate activation andinactivation gating processes. Activation takes place upondepolarization of the membrane (ΔV_(m)) and the channel adopts an openpore conformation allowing Na⁺ influx. Inactivation processes thenchange the channel conformation to a nonconducting, non-activatablestate. Repolarization returns the channels from inactivated to restingconformations. FIG. 2B illustrates how Na⁺ channel opening may beprolonged by toxin binding. Toxins such as veratridine and batrachotoxinare activators that can bind to channels in the open conformation andstabilize the channel in a modified conducting state. This in effectremoves or slows down the inactivation process allowing ion flux tocontinue from minutes to hours. Conversely, toxins such as tetrodotoxin(TTX) are blockers that can bind to the channel in the inactivatedconformations. One method of distinguishing different Na⁺ channels iswhether they are TTX-sensitive or TTX-resistant. (See, for example,Denyer, et al., “HTS Approaches to Voltage-Gated Ion Channel DrugDiscovery”, DDT, 3, No. 7, 323-332 (1998); Whalley, et al. “BasicConcepts in Cellular Cardiac Electrophysiology: Part II: Block of IonChannels by Antiarrhythmic Drugs”, PACE, 18, Part I, 1686-1704 (1995);Goodman & Gilban's “The Pharmacological Basis of Therapeutics”McGraw-Hill, Ninth Ed. Ch. 35, 851-856; and Doggrell, et al., “Ionchannel Modulators as Potential Positive Inotropic Compounds forTreatment of Heart Failure”, Clinical and Experimental Pharmacology andPhysiology, 21833-843, 1994.)

Sodium channel blockers/modulators are employed to alleviate variousdisease conditions including, but not limited to, epilepsy, pain,anaesthesia, neuroprotection, arrhythmia, and migraine. (See, forexample, PCT Publication WO 96/20935, European Patent Application EP0869119, PCT Publication WO 97/27169, U.S. Pat. No. 5,688,830, Hunter &Loughhead “Voltage-Gated Sodium Channel Blockers for the Treatment ofChronic Pain”, Current Opinion in CPNS investigational Drugs, 1999, vol.1, no. 1, 72-81 and Loughhead et al., “Synthesis of MexiletineStereoisomers and Related Compounds via S_(N)Ar NucleophilicSubstitution of a Cr(CO)₈-Complexed Aromatic Fluoride” J. Org. Chem.1999, 64, 3373-3375.) Antiepileptic agents, include, for example,phenytoin, carbamazepine, and lamotrigine. Phenytoin is the prototypicantiepileptic sodium channel blocker and is efficacious in treatingpartial and generalized tonic-clonic seizures in humans. One importantproperty of phenytoin is that it is capable of preventing seizureswithout producing sedation. Thus, phenytoin was the first antiepilepticto approach the therapeutic ideal of inhibiting abnormal brain activitycharacteristic of seizures without appreciably interfering with normalbrain activity.

Carbamazepine, an iminostilbene derivative of tricyclic antidepressants,exhibits a spectrum of anticonvulsant activity very similar to that ofphenytoin. In humans, it is effective against partial and generalizedtonic-clonic seizures, but not against absence seizures. Lamotrigine hasbeen used for treating partial and generalized tonic-clonic seizure.

Topiramate is a sulfamate-substituted monosaccharide, with aphenytoin-like profile in the maximal electroshock and pentylenetetrazoltests. These studies have also shown that it can control seizures insome genetic epilepsy models, in amygdala-kindled rats and in animalswith ischemia-induced epilepsy. Clinical studies have shown thattopiramate is effective as an add-on drug for treating simple or complexpartial seizures with or without secondary generalization, even whenadministrered as monotherapy.

The clinical shortcomings of drugs in current usage are considerable.For example, lamotrigine causes rash and sedation and topiramate,phenytoin, and carbamazepine causes central nervous system side effects.

Thus, there continues to exist a need for novel compounds havingimproved therapeutic activities (e.g., increased potency, greater tissueselectivity, increased efficacy, reduced side effects and a morefavorable duration of action.)

SUMMARY OF THE INVENTION

This invention is directed to novel multibinding compounds that bind toNa⁺ channels in mammalian tissues and can be used to treat diseases andconditions mediated by such channels.

Accordingly, in one of its composition aspects, this invention isdirected to a multibinding compound and salts thereof comprising 2 to 10ligands which may be the same or different and which are covalentlyattached to a linker or linkers, which may be the same or different,each of said ligands comprising a ligand domain capable of binding to aNa⁺ channel.

The multibinding compounds of this invention are preferably representedby formula I:

(L)_(p)(X)_(q)  (I)

where each L is a ligand that may be the same or different at eachoccurrence; X is a linker that may be the same or different at eachoccurrence; p is an integer of from 2 to 10; and q is an integer of from1 to 20; wherein each of said ligands comprises a ligand domain capableof binding to a Na⁺ channel. Preferably q is less than p.

Preferably, the binding of the multibinding compound to a Na⁺ channel orchannels in a mammal modulates diseases and conditions mediated by theNa⁺ channel or channels.

In another of its composition aspects, this invention is directed to apharmaceutical composition comprising a pharmaceutically acceptableexcipient and a therapeutically effective amount of one or moremultibinding compounds (or pharmaceutically acceptable salts thereof)comprising 2 to 10 ligands which may be the same or different and whichare covalently attached to a linker or linkers, which may be the same ordifferent, each of said ligands comprising a ligand domain capable ofbinding to a Na⁺ channel of a cell mediating mammalian diseases orconditions, thereby modulating the diseases or conditions.

In still another of its composition aspects, this invention is directedto a pharmaceutical composition comprising a pharmaceutically acceptableexcipient and a therapeutically effective amount of one or moremultibinding compounds represented by formula I,

(L)_(p)(X)_(q)  (I)

or pharmaceutically acceptable salts thereof, where each L is a ligandthat may be the same or different at each occurrence; X is a linker thatmay be the same or different at each occurrence; p is an integer of from2 to 10; and q is an integer of from 1 to 20; wherein each of saidligands comprises a ligand domain capable of binding to a Na⁺ channel ofa cell mediating mammalian diseases or conditions, thereby modulatingthe diseases or conditions. Preferably q is less than p.

In one of its method aspects this invention is directed to a method formodulating the activity of a Na⁺ channel in a biologic tissue, whichmethod comprises contacting a tissue having a Na⁺ channel with amultibinding compound (or pharmaceutically acceptable salts thereof)under conditions sufficient to produce a change in the activity of thechannel in said tissue, wherein the multibinding compound comprises 2 to10 ligands which may be the same or different and which are covalentlyattached to a linker or linkers, which may be the same or different,each of said ligands comprising a ligand domain capable of binding to aNa⁺ channel.

In another of its method aspects, this invention is directed to a methodfor treating a disease or condition in a mammal resulting from anactivity of a Na⁺ channel, which method comprises administering to saidmammal a therapeutically effective amount of a pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient and oneor more multibinding compounds (or pharmaceutically acceptable saltsthereof) comprising 2 to 10 ligands which may be the same or differentand which are covalently attached to a linker or linkers, which may bethe same or different, each of said ligands comprising a ligand domaincapable of binding to a Na⁺ channel of a cell mediating mammaliandiseases or conditions.

In yet another of its method aspects, this invention is directed to amethod for treating a disease or condition in a mammal resulting from anactivity of a Na⁺ channel, which method comprises administering to saidmammal a therapeutically effective amount of a pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient and oneor more multibinding compounds represented by formula I,

(L)_(p)(X)_(q)  (I)

and pharmaceutically acceptable salts thereof, where each L is a ligandthat may be the same or different at each occurrence; X is a linker thatmay be the same or different at each occurrence; p is an integer of from2 to 10; and q is an integer of from 1 to 20; wherein each of saidligands comprises a ligand domain capable of binding to a Na⁺ channel ofa cell mediating mammalian diseases or conditions. Preferably q is lessthan p.

In a further aspect, this invention provides processes for preparing themultibinding agents of Formula I.

This invention is further directed to general synthetic methods forgenerating large libraries of diverse multimeric compounds whichmultimeric compounds are candidates for possessing multibindingproperties. The diverse multimeric compound libraries provided by thisinvention are synthesized by combining a linker or linkers with a ligandor ligands to provide for a library of multimeric compounds wherein thelinker and ligand each have complementary functional groups permittingcovalent linkage. The library of linkers is preferably selected to havediverse properties such as valency, linker length, linker geometry andrigidity, hydrophilicity or hydrophobicity, amphiphilicity, acidity,basicity and polarization. The library of ligands is preferably selectedto have diverse attachment points on the same ligand, differentfunctional groups at the same site of otherwise the same ligand, and thelike.

This invention is also directed to libraries of diverse emultimericcompounds which multimeric compounds are candidates for possessingmultibinding properties. These libraries are prepared via the methodsdescribed above and permit the rapid and efficient evaluation of whatmolecular constraints impart multibinding properties to a ligand or aclass of ligands targeting a receptor.

Accordingly, in one of its method aspects, this invention is directed toa method for identifying multimeric ligand compounds possessingmultibinding properties which method comprises:

(a) identifying a ligand or a mixture of ligands wherein each ligandcontains at least one reactive functionality;

(b) identifying a library of linkers wherein each linker in said librarycomprises at least two functional groups having complementary reactivityto at least one of the reactive functional groups of the ligand;

(c) preparing a multimeic ligand compound library by combining at leasttwo stoichiometric equivalents of the ligand or mixture of ligandsidentified in (a) with the library of linkers identified in (b) underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands;and

(d) assaying the multimeric ligand compounds produced in (c) above toidentify multimeric ligand compounds possessing multibinding properties.

In another of its method aspects, this invention is directed to a methodfor identifying multimeric ligand compounds possessing multibindingproperties which method comprises:

(a) identifying a library of ligands wherein each ligand contains atleast one reactive functionality;

(b) identifying a linker or mixture of linkers wherein each linkercomprises at least two functional groups having complementary reactivityto at least one of the reactive functional groups of the ligand;

(c) preparing a multimeric ligand compound library by combining at leasttwo stoichiometric equivalents of the library of ligands identified in(a) with the linker or mixture of linkers identified in (b) underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands;and

(d) assaying the multimeric ligand compounds produced in (c) above toidentify multimeric ligand compounds possessing multibinding properties.

The preparation of the multimeric ligand compound library is achieved byeither the sequential or concurrent combination of the two or morestoichiometric equivalents of the ligands identified in (a) with thelinkers identified in (b). Sequential addition is preferred when amixture of different ligands is employed to ensure heterodimeric ormultimeric compounds are prepared. Concurrent addition of the ligandsoccurs when at least a portion of the multimer compounds prepared arehomomultimeric compounds.

The assay protocols recited in (d) can be conducted on the multimericligand compound library produced in (c) above, or preferably, eachmember of the library is isolated by preparative liquid chromatographymass spectrometry (LCMS).

In one of its composition aspects, this invention is directed to alibrary of multimeric ligand compounds which may possess multivalentproperties which library is prepared by the method comprising:

(a) identifying a ligand or a mixture of ligands wherein each ligandcontains at least one reactive functionality;

(b) identifying a library of linkers wherein each linker in said librarycomprises at least two functional groups having complementary reactivityto at least one of the reactive functional groups of the ligand; and

(c) preparing a multimeric ligand compound library by combining at leasttwo stoichiometric equivalents of the ligand or mixture of ligandsidentified in (a) with the library of linkers identified in (b) underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands.

In another of its composition aspects, this invention is directed to alibrary of multimeric ligand compounds which may possess multivalentproperties which library is prepared by the method comprising:

(a) identifying a library of ligands wherein each ligand contains atleast one reactive functionality;

(b) identifying a linker or mixture of linkers wherein each linkercomprises at least two functional groups having complementary reactivityto at least one of the reactive functional groups of the ligand; and

(c) preparing a multimeric ligand compound library by combining at leasttwo stoichiometric equivalents of the library of ligands identified in(a) with the linker or mixture of linkers identified in (b) underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands.

In a preferred embodiment, the library of linkers employed in either themethods or the library aspects of this invention is selected from thegroup comprising flexible linkers, rigid linkers, hydrophobic linkers,hydrophilic linkers, linkers of different geometry, acidic linkers,basic linkers, linkers of different polarization and amphiphiliclinkers. For example, in one embodiment, each of the linkers in thelinker library may comprise linkers of different chain length and/orhaving different complementary reactive groups. Such linker lengths canpreferably range from about 2 to 100 Å.

In another preferred embodiment, the ligand or mixture of ligands isselected to have reactive functionality at different sites on saidligands in order to provide for a range of orientations of said ligandon said multimeric ligand compounds. Such reactive functionalityincludes, by way of example, carboxylic acids, carboxylic acid halides,carboxyl esters, amines, halides, isocyanates, vinyl unsaturation,ketones, aldehydes, thiols, alcohols, anhydrides, and precursorsthereof. It is understood, of course, that the reactive functionality onthe ligand is selected to be complementary to at least one of thereactive groups on the linker so that a covalent linkage can be formedbetween the linker and the ligand.

In other embodiments, the multimeric ligand compound is homomeric (i.e.,each of the ligands is the same, although it may be attached atdifferent points) or heterodimeric (i.e., at least one of the ligands isdifferent from the other ligands).

In addition to the combinatorial methods described herein, thisinvention provides for an interative process for rationally evaluatingwhat molecular constraints impart multibinding properties to a class ofmultimeric compounds or ligands targeting a receptor. Specifically, thismethod aspect is directed to a method for identifying multimeric ligandcompounds possessing multibinding properties which method comprises:

(a) preparing a first collection or iteration of multimeric compoundswhich is prepared by contacting at least two stoichiometric-equivalentsof the ligand or mixture of ligands which target a receptor with alinker or mixture of linkers wherein said ligand or mixture of ligandscomprises at least one reactive functionality and said linker or mixtureof linkers comprises at least two functional groups having complementaryreactivity to at least one of the reactive functional groups of theligand wherein said contacting is conducted under conditions wherein thecomplementary functional groups react to form a covalent linkage betweensaid linker and at least two of said ligands;

(b) assaying said first collection or iteration of multimeric compoundsto assess which if any of said multimeric compounds possess multibindingproperties;

(c) repeating the process of (a) and (b) above until at least onemultimeric compound is found to possess multibinding properties;

(d) evaluating what molecular constraints imparted multibindingproperties to the multimeric compound or compounds found in the firstiteration recited in (a)-(c) above;

(e) creating a second collection or iteration of multimeric compoundswhich elaborates upon the particular molecular constraints impartingmultibinding properties to the multimeric compound or compounds found insaid first iteration;

(f) evaluating what molecular constraints imparted enhanced multibindingproperties to the multimeric compound or compounds found in the secondcollection or iteration recited in (e) above;

(g) optionally repeating steps (e) and (f) to further elaborate uponsaid molecular constraints.

Preferably, steps (e) and (f) are repeated at least two times, morepreferably at from 2-50 times, even more preferably from 3 to 50 times,and still more preferably at least 5-50 times.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are highly schematic illustrations of the transmembraneorganization.

FIGS. 2A and 2B illustrate the multiple ion conducting (open) andnonconducting (closed/inactivated) conformations.

FIG. 3 illustrates a method for optimizing the linker geometry forpresentation of ligands (filled circles) in bivalent compounds:

A. phenyldiacetylene core structure

B. cyclohexane dicarboxylic acid core structure

FIGS. 4A and 4B show exemplary linker “core” structures.

FIG. 5 illustrates examples of multi-binding compounds comprising (A) 2ligands, (B) 3 ligands, (C) 4 ligands, and (D) >4 ligands attached indifferent formats to a linker.

FIG. 6 illustrates a representative ligand which may be used inpreparing multi-binding compounds. Potentially modifiable positions areindicated by arrows.

FIG. 7 illustrates numerous reactive functional groups and the resultingbonds formed by reaction therebetween, and

FIGS. 8A-8Q illustrate convenient methods for preparing the multibindingcompounds of this invention.

DETAILED DESCRIPTION OF THE INVENTION

Biological systems in general are controlled by molecular interactionsbetween bioactive ligands and their receptors, in which the receptor“recognizes” a molecule or a portion thereof (i.e., a ligand domain) toproduce a biological effect. The Na⁺ channels are considered to bepharmacological receptors: they possess specific binding sites forligands having agonist and antagonist activities; the binding of ligandsto such sites modulates Na⁺ flux through the channel; the channelproperties (i.e., gating and ion selectivity) are regulatable.Accordingly, diseases or conditions that involve, or are mediated by,Na⁺ channels can be treated with pharmacologically active ligands thatinteract with such channels to initiate, modulate or abrogatetransporter activity.

The interaction of a Na⁺ channel and a Na⁺ channel-binding ligand may bedescribed in terms of “affinity” and “specificity”; The “affinity” and“specificity” of any given ligand-Na⁺ channel interaction is dependentupon the complementarity of molecular binding surfaces and the energeticcosts of complexation (i.e., the net difference in free energy betweenbound and free states). Affinity may be quantified by the equilibriumconstant of complex formation, the ratio of on/off rate constants,and/or by the free energy of complex formation. Specificity relates tothe difference in binding affinity of a ligand for different receptors.

The net free energy of interaction of such ligand with a Na⁺ channel isthe difference between energetic gains (enthalpy gained throughmolecular complementarity and entropy gained through the hydrophobiceffect) and energetic costs (enthalpy lost through decreased solvationand entropy lost through reduced translational, rotational andconformational degrees of freedom).

The compounds of this invention comprise 2 to 10 Na⁺ channel-bindingligands covalently linked together and capable of acting as multibindingagents. Without wishing to be bound by theory, the enhanced activity ofthese compounds is believed to arise at least in part from their abilityto bind in a multivalent manner with multiple ligand binding sites on aNa⁺ channel or channels, which gives rise to a more favorable net freeenergy of binding. Multivalent interactions differ from collections ofindividual monovalent (univalent) interactions by being capable ofproviding enhanced biologic and/or therapeutic effect. Multivalentbinding can amplify binding affinities and differences in bindingaffinities resulting in enhanced binding specificity as well asaffinity.

DEFINITIONS

As used herein:

The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain, preferably having from 1 to 40 carbonatoms, preferably 1-10 carbon atoms, more preferably 1-6 carbon atoms;such as methyl, ethyl, n-propyl, isopropyl, n-butyl, secondary butyl,tert-butyl, n-hexyl, n-octyl, n-decyl, n-dodecyl, 2-ethyldodecyl,tetradecyl, and the like, unless otherwise indicated.

The term “substituted alkyl” refers to an alkyl group as, defined abovehaving from 1 to 5 substituents selected from the group consisting ofalkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-aryl, —SO₂-heteroaryl, and —NR^(a)R^(b), wherein R^(a)and R^(b) may be the same or different and and are chosen from hydrogen,optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, aryl, heteroaryl and heterocyclic.

The term “alkylene” refers to a diradical of a branched or unbranchedsaturated hydrocarbon chain, preferably having from 1 to 40 carbonatoms, preferably 1-10 carbon atoms, more preferably 1-6 carbon atoms.This term is exemplified by groups such as methylene (—CH₂—), ethylene(—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₃)CH₂—)and the like.

The term “substituted alkylene” refers to: (1) An alkylene group asdefined above having from 1 to 5 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, thioaryloxy,heteroaryl, heteroaryloxy, thioheteroaryloxy, heterocyclic,heterocyclooxy, thioheterocyclooxy, nitro, and —NR_(a)R_(b), whereinR_(a) and R_(b) may be the same or different and are chosen fromhydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. Additionally,such substituted alkylene groups include those where 2 substituents onthe alkylene group are fused to form one or more cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclicor heteroaryl groups fused to the alkylene group; (2) An alkylene groupas defined above that is interrupted by 1-20 atoms independently chosenfrom oxygen, sulfur and NR_(a)—, where R_(a) is chosen from hydrogen,optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic, orgroups selected from carbonyl, carboxyester, carboxyamide and sulfonyl;and (3) An alkylene group as defined above that has both from 1 to 5substituents as defined above and is also interrupted by 1-20 atoms asdefined above. Examples of substituted alkylenes are chloromethylene(—CH(Cl)—), aminoethylene (—CH(NH₂)CH₂—), 2-carboxypropylene isomers(—CH₂CH(CO₂H)CH₂—), ethoxyethyl (—CH₂CH₂O—CH₂CH₂—),ethylmethylaminoethyl (—CH₂CH₂N(CH₃)CH₂CH₂—),1-ethoxy-2-(2-ethoxy-ethoxy)ethane (—CH₂CH₂O—CH₂CH₂—OCH₂CH₂—OCH₂CH₂—),and the like.

The term “alkaryl” or “aralkyl” refers to the groups -alkylene-aryl and-substituted alkylene-aryl in which alkylene and aryl are as definedherein. Such alkaryl groups are exemplified by benzyl, phenethyl and thelike.

The term “alkoxy” refers to the groups alkyl-O—, alkenyl-O—,cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, where alkyl, alkenyl,cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferredalkoxy groups are alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

The term “substituted alkoxy” refers to the groups substituted alkyl-O—,substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein. Examplesof such groups are methylenemethoxy (—CH₂OCH₃), ethylenemethoxy(—CH₂CH₂OCH₃), n-propylene-iso-propoxy (—CH₂CH₂CH₂OCH(CH₃)₂),methylene-t-butoxy (—CH₂—O—C(CH₃)₃) and the like.

The term “alkylthioalkoxy” refers to the group -alkylene-S-alkyl,alkylene-S-substituted alkyl, substituted alkylene-S-alkyl andsubstituted alkylene-S-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylthioakoxy groups are alkylene-S-alkyl and include, by wayof example, methylenethiomethoxy (—CH₂SCH₃), ethylenethiomethoxy(—CH₂CH₂SCH₃), n-propylene-iso-thiopropoxy (—CH₂CH₂CH₂SCH(CH₃)₂),methylene-t-thiobutoxy (—CH₂SC(CH₃)₃) and the like.

“Alkenyl” refers to a monoradical of a branched or unbranchedunsaturated hydrocarbon preferably having from 2 to 40 carbon atoms,preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms, andpreferably having 1-6 double bonds. This term is further exemplified bysuch radicals as vinyl, prop-2-enyl, pent-3-enyl, hex-5-enyl,5-ethlyldodec-3,6-dienyl, and the like.

The term “substituted alkenyl” refers to an alkenyl group as definedabove having from 1 to 5 substituents selected from the group consistingof alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy, substitutedthioalkoxy, aryl, heteroaryl, heterocyclic, aryloxy, thioaryloxy,heteroaryloxy, thioheteroarylosy, heterocyclooxy, thioheterocyclooxy,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, and,—NR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

“Alkenylene” refers to a diradical of an unsaturated hydrocarbon,preferably having from 2 to 40 carbon atoms, preferably 2-10 carbonatoms, more preferably 2-6 carbon atoms, and preferably having 1-6double bonds. This term is further exemplified by such radicals as1,2-ethenyl, 1,3-prop-2-enyl, 1,5-pent-3-enyl, 1,4-hex-5-enyl,5-ethyl-1,12-dodec-3,6-dienyl, and the like.

The term “substituted alkenylene” refers to an alkenylene group asdefined above having from 1 to 5 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano, halogen,hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy,substituted thioalkoxy, aryl, aryloxy, thioaryloxy, heteroaryl,heteroaryloxy, thioheteroaryloxy, heterocyclic, heterocyclooxy,thioheterocyclooxy, nitro, and NR^(a)R^(b), wherein R^(a) and R^(b) maybe the same or different and are chosen from hydrogen, optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,heteroaryl and heterocyclic. Additionally, such substituted alkenylenegroups include those where 2 substituents on the alkenylene group arefused to form one or more cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroarylgroups fused to the alkenylene group.

“Alkynyl” refers to a monoradical of an unsaturated hydrocarbon,preferably having from 2 to 40 carbon atoms, preferably 2-10 carbonatoms, more preferably 2-6 carbon atoms, and preferably having 1-6triple bonds. This term is further exemplified by such radicals asacetylenyl, prop-2-ynyl, pent-3-ynyl, hex-5-ynyl,5-ethyldodec-3,6-diynyl, and the like.

The term “substituted alkenylene” refers to an alkenylene group asdefined above having from 1 to 5 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano, halogen,hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy,substituted thioalkoxy, aryl, aryloxy, thioaryloxy, heteroaryl,heteroaryloxy, thioheteroaryloxy, heterocyclic, heterocyclooxy,thioheterocycloxy, nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, SO₂-heterocyclic, NR^(a)R^(b), wherein R^(a) and R^(b)may be the same or different and are chosen from hydrogen, optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,heteroaryl and heterocyclic.

“Alkynylene” refers to a diradical of an unsaturated hydrocarbonradical, preferably having from 2 to 40 carbon atoms, preferably 2-10carbon atoms, more preferably 2-6 carbon atoms, and preferably having1-6 triple bonds. This term is further exemplified by such radicals as1,3-prop-2-ynyl, 1,5-pent-3-ynyl, 1,4-hex-5-ynyl,5-ethyl-1,12-dodec-3,6-diynyl, and the like.

The term “acyl” refers to the groups —CHO, alkyl-C(O)—, substitutedalkyl-C(O)—, cycloalkyl-C(O)—, substituted cycloalkyl-C(O)—,cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—, aryl-C(O)—,heteroaryl-C(O)— and heterocyclic-C(O)— where alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.

The term “acylamino” refers to the group —C(O)NRR where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,heterocyclic or where both R groups are joined to form a heterocyclicgroup (e.g., morpholine) wherein alkyl, substituted alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

The term “aminoacyl” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

The term “aminoacyloxy” refers to the group —NRC(O)OR where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclic-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclic are as defined herein.

The term “aryl” refers to an unsaturated aromatic carbocyclic group offrom 6 to 20 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed (fused) rings (e.g., naphthyl or anthryl).

Unless otherwise constrained by the definition for the aryl substituent,such aryl groups can optionally be substituted with from 1 to 5substituents selected from the group consisting of acyloxy, hydroxy,thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,substituted alkyl, substituted alkoxy, substituted alkenyl, substitutedalkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino,aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl,carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —S₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, trihalomethyl,NR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy.

The term “aryloxy” refers to the group aryl-O— wherein the aryl group isas defined above including optionally substituted aryl groups as alsodefined above.

The term “arylene” refers to a diradical derived from aryl orsubstituted aryl as defined above, and is exemplified by 1,2-phenylene,1,3-phenylene, 1,4-phenylene, 1,2-naphthylene and the like.

The term “amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl and heterocyclic provided thatboth R's are not hydrogen.

The term “carboxyalkyl” refers to the group “—C(O)O-alkyl”,“—C(O)O-substituted alkyl”, “—C(O)O-cycloalkyl”, “—C(O)O-substitutedcycloalkyl”, “—C(O)O-alkenyl”, “—C(O)O-substituted alkenyl”,“—C(O)O-alkynyl” and “—C(O)O-substituted alkynyl” where alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl and substituted alkynyl where alkynyl areas defined herein.

The term “cycloalkyl” refers to cyclic alkyl groups of from 3 to 20carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups havingfrom 1 to 5 substituents selected from the group consisting of alkoxy,substituted alkoxy, cycloalkyl, cycloalkenyl, substituted cycloalkenyl,acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl,azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, andNR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

The term “cycloalkenyl” refers to cyclic alkenyl groups of from 4 to 20carbon atoms having a single cyclic ring or fused rings and at least onepoint of internal unsaturation. Examples of suitable cycloalkenyl groupsinclude, for instance, cyclobut-2-enyl, cyclopent-3-enyl,cyclooct-3-enyl and the like.

The term “substituted cycloalkenyl” refers to cycloalkenyl groups havingfrom 1 to 5 substituents selected from the group consisting of alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl,aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto,thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl;—SO₂-heteroaryl, and NR^(a)R^(b), wherein R^(a) and R^(b) may be thesame or different and are chosen from hydrogen, optionally substitutedalkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl andheterocyclic.

The term “halo” or “halogen” refers to fluoro, chloro, bromo and iodo.

“Haloalkyl” refers to alkyl as defined above substituted by 1-4 halogroups as defined above, which may be the same or different, such as3-fluorododecyl, 12,12,12-trifluorododecyl, 2-bromooctyl,-3-bromo-6-chloroheptyl, and the like.

The term “heteroaryl” refers to an aromatic group of from 1 to 15 carbonatoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfurwithin at least one ring (if there is more than one ring).

Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 5 substituents selected from the group consisting of acyloxy,hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, substituted alkyl, substituted alkoxy, substitutedalkenyl, substituted alkynyl, substituted cycloalkyl, substitutedcycloalkenyl, amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy,azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, trihalomethyl,mono-and di-alkylamino, mono- and NR^(a)R^(b), wherein R^(a) and R^(b)may be the same or different and are chosen from hydrogen, optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,heteroaryl and heterocyclic. Preferred heteroaryls include pyridyl,pyrrolyl and furyl.

The term “heteroaryloxy” refers to the group heteroaryl-O—.

The term “heteroarylene” refers to the diradical group derived fromheteroaryl or substituted heteroaryl as defined above, and isexemplified by the groups 2,6-pyridylene, 2,4-pyridiylene,1,2-quinolinylene, 1,8-quinolinylene, 1,4-benzofuranylene,2,5-pyridiylene, 1,3-morpholinylene, 2,5-indolenyl and the like.

The term “heterocycle” or “heterocyclic” refers to a monoradicalsaturated or unsaturated group having a single ring or multiplecondensed rings, from 1 to 40 carbon atoms and from 1 to 10 heteroatoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 5, and preferably 1 to 3 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, cyano; halogen,hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, and NR^(a)R^(b),wherein R^(a) and R^(b) may be the same or different and are chosen fromhydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. Suchheterocyclic groups can have a single ring or multiple condensed rings.

Examples of nitrogen heterocycles and heteroaryls include, but are notlimited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridie,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, incline, morpholino, piperidinyl,tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containingheterocycles.

A preferred class of heterocyclics include “crown compounds” whichrefers to a specific class of heterocyclic compounds having one or morerepeating units of the formula [—(CH₂—)_(m)Y—] where m is equal to orgreater than 2, and Y at each separate occurrence can be O, N, S or P.Examples of crown compounds include, by way of example only,[—(CH₂)₃—NH—]₃, [—((CH₂)₂—O)₄—((CH₂)₂—NH)₂] and the like. Typically suchcrown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbonatoms.

The term “heterocyclooxy” refers to the group heterocyclic-O—.

The term “thioheterocyclooxy” refers to the group heterocyclic-S—.

The term “heterocyclene” refers to the diradical group derived from aheterocycle as defined herein, and is exemplified by the groups2,6-morpholino, 2,5-morpholino and the like.

The term “oxyacylamino” refers to the group —OC(O)NRR where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

The term “thiol” refers to the group —SH.

The term “thioalkoxy” refers to the group —S-alkyl.

The term “substituted thioalkoxy” refers to the group —S-substitutedalkyl

The term “thioaryloxy” refers to the group aryl-S— wherein the arylgroup is as defined above including optionally substituted aryl groupsalso defined above.

The term “thioheteroaryloxy” refers to the group heteroaryl-S— whereinthe heteroaryl group is as defined above including optionallysubstituted aryl groups as also defined above.

As to any of the above groups which contain one or more substituents, itis understood, of course, that such groups do not contain anysubstitution or substitution patterns which are sterically impracticaland/or synthetically non-feasible. In addition, the compounds of thisinvention include all stereochemical isomers arising from thesubstitution of these compounds.

“Alkyl optionally interrupted by 1-5 atoms chosen from O, S, or N”refers to alkyl as defined above in which the carbon chain isinterrupted by O, S, or N. Within the scope are ethers, sulfides, andamines, for example 1-methoxydecyl, 1-pentyloxynonane,1-(2-isopropoxyethoxy)-4-methylnonane, 1-(2-ethoxyethoxy)dodecyl,2-(t-butoxy)heptyl, 1-pentylsulfanylnonane, nonylpentylamine, and thelike.

“Heteroarylalkyl” refers to heteroaryl as defined above linked to alkylas defined above, for example pyrid-2-ylmethyl, 8-quinolinylpropyl, andthe like.

“Optional” or “optionally” means that the subsequently described eventor circumstance may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, optionally substituted alkyl means that alkylmay or may not be substituted by those groups enumerated in thedefinition of substituted alkyl.

The term “pharmaceutically acceptable salt” refers to salts which retainthe biological effectiveness and properties of the multibindingcompounds of this invention and which are not biologically or otherwiseundesirable. In many cases, the multibinding compounds of this inventionare capable of forming acid and/or base salts by virtue of the presenceof amino and/or carboxyl groups or groups similar thereto.

Pharmaceutically acceptable base addition salts can be prepared frominorganic and organic bases. Salts derived from inorganic bases, includeby way of example only, sodium, potassium, lithium, ammonium, calciumand magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amities, trialkenyl amines, substituted alkenylamines, di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group.

Examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine,tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike. It should also be understood that other carboxylic acidderivatives would be useful in the practice of this invention, forexample, carboxylic acid amides, including carboxamides, lower alkylcarboxamides, dialkyl carboxamides, and the like.

Pharmaceutically acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid,salicylic acid, and the like.

The term “protecting group” or “blocking group” refers to any groupwhich when bound to one or more hydroxyl, thiol, amino or carboxylgroups of the compounds prevents reactions from occurring at thesegroups and which protecting group can be removed by conventionalchemical or enzymatic steps to reestablish the hydroxyl, thiol, amino orcarboxyl group. See, generally, T. W. Greene & P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 2^(nd) Ed., 1991, John Wiley and Sons, N.Y.

The particular removable blocking group employed is not critical andpreferred removable hydroxyl blocking groups include conventionalsubstituents such as allyl, benzyl, acetyl, chloroacetyl, thiobenzyl,benzylidine, phenacyl, t-butyl-diphenylsilyl and any other group thatcan be introduced chemically onto a hydroxyl functionality and laterselectively removed either by chemical or enzymatic methods in mildconditions compatible with the nature of the product.

Preferred removable amino blocking groups include conventionalsubstituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ),fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like,which can be removed by conventional conditions compatible with thenature of the product.

Preferred carboxyl protecting groups include esters such as methyl,ethyl propyl, t-butyl etc. which can be removed by mild hydrolysisconditions compatible with the nature of the product.

As used herein, the terms “inert organic solvent” or “inert solvent”mean a solvent inert under the conditions of the reaction beingdescribed in conjunction therewith including, for example, benzene,toluene, acetonitrile, tetrahydrofuran, dimethylformamide, chloroform,methylene chloride (or dichloromethane), diethyl ether, ethyl acetate,acetone, methylethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

The term “Na⁺ channel” refers to a structure comprised of integralmembrane proteins that functions to allow Na⁺ to equilibrate across amembrane according to its electrochemical gradient and at rates that arediffusion limited.

“Ligand” as used herein denotes a compound that is a binding partner fora Na⁺ channel receptor, and is bound thereto, for example, bycomplementarity. The specific region or regions of the ligand moleculethat is recognized by the ligand binding site of a Na⁺ channel receptoris designated as the “ligand domain”. A ligand may be either capable ofbinding to a receptor by itself, or may require the presence of one ormore non-ligand components for binding (e.g. ions, a lipid molecule, asolvent molecule, and the like). The linker can be either a chiral orachiral molecule.

The ligands and linkers which comprise the multibinding agents of theinvention and the multibinding compounds themselves may have varioussteroisomeric forms, including enantiomers and diastereomers. It is tobe understood that the invention contemplates all possiblestereoisomeric forms of multibinding compounds, and mixtures thereof.

Ligands useful in this invention comprise Na⁺ channel modulators suchas, for example, carbamazepine, felbamate, fosphenytoin, lamotrigine,permenol, topiramate, vipocitine, phenytoin, ADC1, alprafenone, trophix,AWD-140-190, berlafenone, BRB-I-28, CI-953, CNS-5151, Co-102862,E-047/1, GE-68, GW273227, GW286103, GW273293, iodoamiloride, lidocaine,PNU-151774E, PD-85639, RP-66055, RSD-921, RS-2135, SL-90.0571,sipatrigine, topiramate, QX-314, ZM-227189, 534U87, 4030W92, 202W92mexilitene, N-ethylmexilitene, flecainide, RS 132943, and tocainide andtheir analogues. Table 1 sets forth the indications treated by the Na⁺channel modulators. It should be noted that beyond the primaryindications listed in the table, many of the Na⁺ channel blockers suchas, for example, mexilitene, lamotrigine, amitriptyline, and otheranti-seizure compounds are used to treat pain as well.

TABLE 1 Drug Indication(s) Carbamazepine Epilepsy Felbamate EpilepsyFosphenytoin Epilepsy Lamotrigine Epilepsy, ischemia, seizures PermenolArrythmia Topiramate Epilepsy, seizures Vipocitine Epilepsy, depressionPhenytoin Seizures ADC1 Ischemia, epilepsy, seizure AlprafenoneArrhythmia Trophix Pain AWD-140-190 Epilepsy, CNS disease BerlafenoneArrhythmia BRB-I-28 Arrhythmia CI-953 Ischemia CNS-5151 IschemiaCo-102862 Pain, epilepsy E-047/1 Arrythmia GE-68 Arrythmia GW273227GW286103 GW273293 Epilepsy, pain Iodoamiloride Cystic fibrosis LidocaineArrhythmia PNU-151774E Pain, epilepsy PD-85639 Arrythmia RP-66055Cerebral infarction, cerebrovascular ischemia, epilepsy RSD-921Arrhythmia RS-2135 Arrhythmia SL-90.0571 Epilepsy Sipatrigine StrokeTopiramate Epilepsy (children) QX-314 pain/urinary tract diseaseZM-227189 Arrythmia, tachycardia 534U87 Epilepsy 4030W92 Epilepsy, pain,depression 202W92 Stroke mexilitene Pain N-ethylmexilitene FlecainidePain RS 132943 Pain Tocainide Amitriptyline Pain

While it is contemplated that many sodium channel ligands that arecurrently known can be used in the preparation of multibinding compoundsof this invention, it should be understood that portions of the ligandstructure that are not essential for molecular recognition and bindingactivity (i.e., that are not part of the ligand domain) may be variedsubstantially, replaced with unrelated structures and, in some cases,omitted entirely without affecting the binding interaction. Accordingly,it should be understood that the term “ligand” is not intended to belimited to compounds known to be useful as Na⁺ channel receptor-bindingcompounds (e.g., known drugs), in that ligands that exhibit marginalactivity or lack useful activity as monomers can be highly active asmultibinding compounds, because of the biological benefit conferred bymultivalency. The primary requirement for a ligand as defined herein isthat it has a ligand domain, as defined above, which is available forbinding to a recognition site on a Na⁺ channel.

For purposes of the present invention, the term “ligand” or “legands” isintended to include the racemic ligands as well as the individualstereoisomers of the ligands, including pure enantiomers andnon-raceunic mixtures thereof. The scope of the invention as describedand claimed encompasses the racemic forms of the ligands as well as theindividual enantiomers and non-racemic mixtures thereof.

The term “ligand binding site” as used herein denotes a site on a Na⁺channel receptor that recognizes a ligand domain and provides a bindingpartner for the ligand. The ligand binding site may be defined bymonomeric or multimeric structures. This interaction may be capable ofproducing a unique biological effect, for example agonism, antagonism,modulation, or may maintain an ongoing biological event, and the like.

It should be recognized that the ligand binding sites of Na⁺ channelreceptors that participate in biological multivalent bindinginteractions are constrained to varying degrees by their intra- andintermolecular associations. For example, Na⁺ channel ligand bindingsites may be covalently joined in a single structure, noncovalentlyassociated in one or more multimeric structures, embedded in a membraneor biopolymer matrix, and so on, and therefore have less translationaland rotational freedom than if the same sites were present as monomersin solution.

The terms “agonism” and “antagonism” are well known in the art. As usedherein, the term “agonist” refers to a ligand that when bound to a Na⁺channel stimulates its activity. The term “antagonist” refers to aligand that when bound to a Na⁺ channel inhibits its activity. Channelblock or activation may result from allosteric effects of ligand bindingto the channel rather than occupancy of the channel pore. Theseallosteric effects may produce changes in protein conformation thataffect Na⁺ binding sites, gating mechanisms and/or the pore region(i.e., ion permeation).

As described above, a sodium channel can exist in several modes: C(closed resting state); C* (activated closed state); O (open state); andI (inactivated state). The probability that a channel will exist in oneof these four states changes with voltage. A given ligand may havedifferent binding affinities for different states, and be capable ofproducing agonist or antagonist activity.

The term “modulatory effect” is intended to refer to the ability of aligand to change the activity of a Na⁺ channel through binding to thechannel.

“Multibinding agent” or “multibinding compound” refers herein to acompound that has from 2 to 10 Na⁺ channel ligands as defined herein(which may be the same or different) covalently bound to one or morelinkers (which may be the same or different), and is capable ofmultivalency, as defined below.

A multibinding compound provides an improved biologic an d/ortherapeutic effect compared to that of the same number of unlinkedligands available for binding to the ligand binding sites on a Na⁺channel or channels. Examples of improved “biologic and/or therapeuticeffect” include increased ligand-receptor binding interactions (e.g.,increased affinity, increased ability to elicit a functional change inthe target, improved kinetics), increased selectivity for the target,increased potency, increased efficacy, decreased toxicity, increasedtherapeutic index, improved duration of action, improvedbioavailability, improved pharmacokinetics, improved activity spectrum,and the like. The multibinding compounds of this invention will exhibitat least one, and preferably more than one, of the above-mentionedeffects.

The term “library” refers to at least 3, preferably from 10² to 10⁹ andmore preferably from 10² to 10⁴ multimeric compounds. Preferably, thesecompounds are prepared as a multiplicity of compounds in a singlesolution or reaction mixture which permits facile synthesis thereof. Inone embodiment, the library of multimeric compounds can be directlyassayed for multibinding properties. In another embodiment, each memberof the library of multimeric compounds is first isolated and,optionally, characterized. This member is then assayed for multibindingproperties.

The term “collection” refers to a set of multimeric compounds which areprepared either sequentially or concurrently (e.g., combinatorially).The collection comprises at least 2 members; preferably from 2 to 10⁹members and still more preferably from 10 to 10⁴ members.

The term “multimeric compound” refers to compounds comprising from 2 to10 ligands covalently connected through at least one linker whichcompounds may or may not possess multibinding properties (as definedherein).

The term “pseudohalide” refers to functional groups which react indisplacement reactions in a manner similar to a halogen. Such functionalgroups include, by way of example; mesyl, tosyl, azido and cyano groups.

“Univalency” or “monovalency” as used herein refers to a single bindinginteraction between one ligand with one ligand binding site as definedherein. It should be noted that a compound having multiple copies of aligand (or ligands) exhibits univalency when only one ligand of thatcompound interacts with a ligand binding site. Examples of univalentinteractions are depicted below.

“Multivalency” as used herein refers to the concurrent binding of from 2to 10 linked ligands, which may be the same or different, and two ormore corresponding ligand binding sites; which may be the same ordifferent. An example of trivalent binding is depicted below forillustrative purposes.

It should be understood that not all compounds that contain multiplecopies of a ligand attached to a linker necessarily exhibit thephenomena of multivalency, i.e., that the biologic and/or therapeuticeffect of the multibinding agent is greater than that of the same numberof unlinked ligands made available for binding to the ligand bindingsites. For multivalency to occur, the ligand domains of the ligands thatare linked together must be presented to their cognate ligand bindingsites by the linker or linkers in a specific manner in order to bringabout the desired ligand-orienting result, and thus produce amultibinding interaction.

The term “linker” or “linkers” as used herein, identified whereappropriate by the symbol X, refers to a group or groups that covalentlylink(s) from 2 to 10 ligands (as defined above) in a manner thatprovides a compound capable of multivalency. The linker is aligand-orienting entity that permits attachment of multiple copies of aligand (which may be the same or different) thereto.

The term “linker” includes everything that is not considered to be partof the ligand, e.g., ancillary groups such as solubilizing groups,lipophilic groups, groups that alter pharmacodynamics orpharmacokinetics, groups that modify the diffusability of themultibinding compound, spacers that attach the ligand to the linker,groups that aid the ligand-orienting function of the linker, forexample, by imparting flexibility or rigidity to the linker as a whole,or to a portion thereof, and so on. The term “linker” does not, however,cover solid inert supports such as beads, glass particles, rods, and thelike, but it is to be understood that the multibinding compounds of thisinvention can be attached to a solid support if desired, for example,for use in separation and purification processes and for similarapplications.

The extent to which the previously discussed enhanced activity ofmultibinding compounds is realized in this invention depends upon theefficiency with which the linker or linkers that joins the ligandspresents them to their array of ligand binding sites. Beyond presentingthese ligands for multivalent interactions with ligand binding sites,the linker spatially constrains these interactions to occur withindimensions defined by the linker.

The linkers used in this invention are selected to allow multivalentbinding of ligands to any desired ligand binding sites of a Na⁺ channel,whether such sites are located within the cell membrane, interiorly(e.g., within a channel/translocation pore), both interiorly and on theperiphery of a channel, at the boundary region between the lipid bilayerand the channel, or at any intermediate position thereof. The preferredlinker length will vary depending on the distance between adjacentligand binding sites, and the geometry, flexibility and composition ofthe linker. The length of the linker will preferably be in the range ofabout 2 Å to about 100 Å, more preferably from about 2 Å to about 50 Åand even more preferably from about 3 Å to about 20 Å.

The ligands are covalently attached to the linker or linkers usingconventional chemical techniques. The reaction chemistries resulting insuch linkage are well known in the art and involve the use of reactivefunctional groups present on the linker and ligand. Preferably, thereactive functional groups on the linker are selected relative to thefunctional groups available on the ligand for coupling, or which can beintroduced onto the ligand for this purpose. Again, such reactivefunctional groups are well known in the art. For example, reactionbetween a carboxylic acid of either the linker or the ligand and aprimary or secondary amine of the ligand or the linker in the presenceof suitable well-known activating agents results in formation of anamide bond covalently linking the ligand to the linker; reaction betweenan amine group of either the linker or the ligand and a sulfonyl halideof the ligand or the linker results in formation of a sulfonamide bondcovalently linking the ligand to the linker; and reaction between analcohol or phenol group of either the linker or the ligand and an alkylor aryl halide of the ligand or the linker results in formation of anether bond covalently linking the ligand to the linker. FIG. 7illustrates numerous reactive functional groups and the resulting bondsformed by reaction therebetween. Where functional groups are lacking,they can be created by suitable chemistries that are described instandard organic chemistry texts such as J. March, Advanced OrganicChemistry, 4^(th) Ed., (Wiley-Interscience, N.Y., 1992).

The linker is attached to the ligand at a position that retains liganddomain-ligand binding site interaction and specifically which permitsthe ligand domain of the ligand to orient itself to bind to the ligandbinding site. Such positions and synthetic protocols for linkage arewell known in the art. The term linker embraces everything that is notconsidered to be part of the ligand.

The relative orientation in which the ligand domains are displayeddepends both on the particular point or points of attachment of theligands to the linker, and on the framework geometry. The determinationof where acceptable substitutions can be made on a ligand is typicallybased on prior knowledge of structure-activity relationships (SAR) ofthe ligand and/or congeners and/or structural information aboutligand-receptor complexes (e.g. X-ray crystallography, NMR, and thelike). Such positions and synthetic protocols for linkage are well knownin the art and can be determined by those with ordinary skill in the art(see Methods of Preparation.) Following attachment of a ligand to thelinker or linkers, or to a significant portion thereof (e.g., 2-10 atomsof linker), the linker-ligand conjugate may be tested for retention ofactivity in a relevant assay system (see Utility and Testing below forrepresentative assays).

At present, it is preferred that the multibinding compound is a bivalentcompound in which two ligands are covalently linked, or a trivalentcompound, in which three ligands are covalently linked. Linker design isfurther discussed under Methods of Preparation.

“Potency” as used herein refers to the minimum concentration at which aligand is able to achieve a desirable biological or therapeutic effect.The potency of a ligand is typically proportional to its affinity forits receptor. In some cases, the potency may be non-linearly correlatedwith its affinity. In comparing the potency of two drugs, e.g., amultibinding agent and the aggregate of its unlinked ligand, thedose-response curve of each is determined under identical testconditions (e.g., in an in vitro or in vivo assay, in an appropriateanimal model). The finding that the multibinding agent produces anequivalent biologic or therapeutic effect at a lower concentration thanthe aggregate unlinked ligand (e.g., on a per weight, per mole or perligand basis) is indicative of enhanced potency.

“Selectivity” or “specificity” is a measure of the binding preferencesof a ligand for different receptors. The selectivity of a ligand withrespect to its target receptor relative to another receptor is given bythe ratio of the respective values of K_(d) (i.e., the dissociationconstants for each ligand-receptor complex) or, in cases where abiological effect is observed below the K_(d), the ratio of therespective EC₅₀s or IC₅₀s (i.e., the concentrations that produce 50% ofthe maximum response for the ligand interacting with the two distinctreceptors).

The term “treatment” refers to any treatment of a disease or conditionin a mammal, particularly a human, and includes:

(i) preventing the disease or condition from occurring in a subjectwhich may be predisposed to the condition but has not yet been diagnosedwith the condition and, accordingly, the treatment constitutesprophylactic treatment for the pathologic condition;

(ii) inhibiting the disease or condition, i.e., arresting itsdevelopment;

(iii) relieving the disease or condition, i.e., causing regression ofthe disease or condition; or

(iv) relieving the symptoms resulting from the disease or conditionwithout addressing the underlying disease or condition, e.g., relievingsymptoms of epilepsy, seizures, pain, stroke, ischemia, arrhythmia anddepression, but not an underlying cause.

The phrase “disease or condition which is modulated by treatment with amultibinding Na⁺ channel ligand” covers all disease states and/orconditions that are generally acknowledged in the art to be usefullytreated with a ligand for a Na⁺ channel in general, and those diseasestates and/or conditions that have been found to be usefully treated bya specific multibinding compound of our invention, i.e., the compoundsof Formula I. Such disease states include, by way of example only,pathophysiological disorders, including hypertension, cardiacarrhythmogenesis, insulin-dependent diabetes, non-insulin dependentdiabetes mellitus, diabetic neuropathy, seizures, tachycardia, ischemicheart disease, cardiac failure, angina, myocardial infarction,transplant rejection, autoimmune disease, sickle cell anemia, musculardystrophy, gastrointestinal disease, mental disorder, sleep disorder,anxiety disorder, eating disorder, neurosis, alcoholism, inflammation,cerebrovascular ischemia, CNS diseases, epilepsy, Parkinson's disease,asthma, incontinence, urinary dysfunction, micturition disorder,irritable bowel syndrome, restenosis, subarachnoid hemorrhage,Alzheimers disease, drug dependence/addiction, schizophrenia,Huntington's chorea, tension-type headache, trigeminal neuralgia,cluster headache, migraine (acute and prophylaxis), inflammatory pain,neuropathic pain and depression.

The term “therapeutically effective amount” refers to that amount ofmultibinding compound that is sufficient to effect treatment, as definedabove, when administered to a mammal in need of such treatment. Thetherapeutically effective amount will vary depending upon the subjectand disease condition being treated, the weight and age of the subject,the severity of the disease condition, the manner of administration andthe like, which can readily be determined by one of ordinary skill inthe art.

The term “pharmaceutically acceptable excipient” is intended to includevehicles and carriers capable of being coadministered with amultibinding compound to facilitate the performance of its intendedfunction. The use of such media for pharmaceutically active substancesis well known in the art. Examples of such vehicles and carriers includesolutions, solvents, dispersion media, delay agents, emulsions and thelike. Any other conventional carrier suitable for use with themultibinding compounds also falls within the scope of the presentinvention.

Combinatorial Libraries

The methods described above lend themselves to combinatorial approachesfor identifying multimeric compounds which possess multibindingproperties. Specifically, factors such as the proper juxtaposition ofthe individual ligands of a multibinding compound with respect to therelevant array of binding sites on a target or targets is important, inoptimizing the interaction of the multibinding compound with itstarget(s) and to maximize the biological advantage through multivalency.One approach is to identify a library of candidate multibindingcompounds with properties spanning the multibinding parameters that arerelevant for a particular target. These parameters include: (1) theidentity of ligand(s), (2) the orientation of ligands, (3) the valencyof the construct, (4) linker length, (5) linker geometry, (6) linkerphysical properties, and (7) linker chemical functional groups.

Libraries of multimeric compounds potentially possessing multibindingproperties (i.e., candidate multibinding compounds) and comprising amultiplicity of such variables are prepared and these libraries are thenevaluated via conventional assays corresponding to the ligand selectedand the multibinding parameters desired. Considerations relevant to eachof these variables are set forth below:

Selection of ligand(s)

A single ligand or set of ligands is (are) selected for incorporationinto the libraries of candidate multibinding compounds which library isdirected against a particular biological target or targets. The onlyrequirement for the ligands chosen is that they are capable ofinteracting with the selected target(s). Thus, ligands may be knowndrugs, modified forms of known drugs, substructures of known drugs orsubstrates of modified forms of known drugs (which are competent tointeract with the target), or other compounds. Ligands are preferablychosen based on known favorable properties that may be projected to becarried over to or amplified in multibinding forms. Favorable propertiesinclude demonstrated safety and efficacy in human patients, appropriatePK/ADME profiles, synthetic accessibility, and desirable physicalproperties such as solubility, logP, etc. However, it is crucial to notethat ligands which display an unfavorable property from among theprevious list may obtain a more favorable property through the processof multibinding compound formation; i.e., ligands should not necessarilybe excluded on such a basis. For example, a ligand that is notsufficiently potent at a particular target so as to be efficacious in ahuman patient may become highly potent and efficacious when presented inmultibinding form. A ligand that is potent and efficacious but not ofutility because of a non-mechanism-related toxic side effect may haveincreased therapeutic index (increased potency relative to toxicity) asa multibinding compound. Compounds that exhibit short in vivo half-livesmay have extended half-lives as multibinding compounds. Physicalproperties of ligands that limit their usefulness (e.g. poorbioavailability due to low solubility, hydrophobicity, hydrophilicity)may be rationally modulated in multibinding forms, providing compoundswith physical properties consistent with the desired utility.

Orientation: selection of ligand attachment points and linking chemistry

Several points are chosen on each ligand at which to attach the ligandto the linker. The selected points on the ligand/linker for attachmentare functionalized to contain complementary reactive functional groups.This permits probing the effects of presenting the ligands to theirreceptor(s) in multiple relative orientations, an important multibindingdesign parameter. The only requirement for choosing attachment points isthat attaching to at least one of these points does not abrogateactivity of the ligand. Such points for attachment can be identified bystructural information when available. For example, inspection of aco-crystal structure of a protease inhibitor bound to its target allowsone to identify one or more sites where linker attachment will notpreclude the enzyme:inhibitor interaction. Alternatively, evaluation ofligand/target binding by nuclear magnetic resonance will permit theidentification of sites non-essential for ligand/target binding. See,for example, Fesik, et al., U.S. Pat. No. 5,891,643. When suchstructural information is not available, utilization ofstructure-activity relationships (SAR) for ligands will suggestpositions where substantial structural variations are and are notallowed. In the absence of both structural and SAR information, alibrary is merely selected with multiple points of attachment to allowpresentation of the ligand in multiple distinct orientations. Subsequentevaluation of this library will indicate what positions are suitable forattachment.

It is important to emphasize that positions of attachment that doabrogate the activity of the monomeric ligand may also be advantageouslyincluded in candidate multibinding compounds in the library providedthat such compounds bear at least one ligand attached in a manner whichdoes not abrogate intrinsic activity. This selection derives from, forexample, heterobivalent interactions within the context of a singletarget molecule. For example, consider a receptor antagonist ligandbound to its target receptor, and then consider modifying this ligand byattaching to it a second copy of the same ligand with a linker whichallows the second ligand to interact with the same receptor molecule atsites proximal to the antagonist binding site, which include elements ofthe receptor that are not part of the formal antagonist binding siteand/or are elements of the matrix surrounding the receptor such as themembrane. Here, the most favorable orientation for interaction of thesecond ligand molecule with the receptor/matrix may be achieved byattaching it to the linker at a position which abrogates activity of theligand at the formal antagonist binding site. Another way to considerthis is that the SAR of individual ligands within the context of amultibinding structure is often different from the SAR of those sameligands in momomeric form.

The foregoing discussion focused on bivalent interactions of dimericcompounds bearing two copies of the same ligand joined to a singlelinker through different attachment points, one of which may abrogatethe binding/activity of the monomeric ligand. It should also beunderstood that bivalent advantage may also be attained withheterodimeric constructs bearing two different ligands that bind tocommon or different targets.

For example, an Na⁺ channel blocker and an opioid agonist may be joinedto a linker through attachment points which do not abrogate the bindingaffinity of the monomeric ligands for their respective receptor sites.Both target receptors are present on CNS nerve cells. If the opioidagonist unit enhances the activity of Na⁺ channel blocker at the mostimportant Na⁺ channels, and the Na⁺ channel blocker enhances theactivity of the opioid agonist at the appropriate opioid receptors, theactivity will be above and beyond that of the combination of themonomeric species.

Once the ligand attachment points have been chosen, one identifies thetypes of chemical linkages that are possible at those points. The mostpreferred types of chemical linkages are those that are compatible withthe overall structure of the ligand (or protected forms of the ligand)readily and generally formed, stable and intrinsically inocuous undertypical chemical and physiological conditions, and compatible with alarge number of available linkers. Amide bonds, ethers, amines,carbamates, ureas, and sulfonamides are but a few examples of preferredlinkages.

Linkers: spanning relevant multibinding parameters through selection ofvalency, linker length, linker geometry, rigidity, physical properties,and chemical functional groups

In the library of linkers employed to generate the library of candidatemultibinding compounds, the selection of linkers employed in thislibrary of linkers takes into consideration the following factors:

Valency. In most instances the library of linkers is initiated withdivalent linkers. The choice of ligands and proper juxtaposition of twoligands relative to their binding sites permits such molecules toexhibit target binding affinities and specificities more than sufficientto confer biological advantage. Furthermore, divalent linkers orconstructs are also typically of modest size such that they retain thedesirable biodistribution properties of small molecules.

Linker length. Linkers are chosen in a range of lengths to allow thespanning of a range of inter-ligand distances that encompass thedistance preferable for a given divalent interaction. In some instancesthe preferred distance can be estimated rather precisely fromhigh-resolution structural information of targets, typically enzymes andsoluble receptor targets. In other instances where high-resolutionstructural information is not available (such as 7TM G-protein coupledreceptors), one can make use of simple models to estimate the maximumdistance between binding sites either on adjacent receptors or atdifferent locations on the same receptor. In situations where twobinding sites are present on the same target (or target subunit formultisubunit targets), preferred linker distarices are 2-20 Å, with morepreferred linker distances of 3-12 Å. In situations where two bindingsites reside on separate (e.g., protein) target sites, preferred linkerdistances are 20-100 Å, with more preferred distances of 30-70 Å.

Linker geometry and rigidity. The combination of ligand attachment site,linker length, linker geometry, and linker rigidity determine thepossible ways in which the ligands of candidate multibinding compoundsmay be displayed in three dimensions and thereby presented to theirbinding sites. Linker geometry and rigidity are nominally determined bychemical composition and bonding pattern, which may be controlled andare systematically varied as another spanning function in a multibindingarray. For example, linker geometry is varied by attaching two ligandsto the ortho, meta, and para positions of a benzene ring, or in cis- ortrans-arrangements at the 1,1- vs. 1,2- vs. 1,3- vs. 1,4-positionsaround a cyclohexane core or in cis- or trans-arrangements at a point ofethylene unsaturation. Linker rigidity is varied by controlling thenumber and relative energies of different conformational states possiblefor the linker. For example, a divalent compound bearing two ligandsjoined by 1,8-octyl linker has many more degrees of freedom, and istherefore less rigid than a compound in which the two ligands areattached to the 4,4′ positions of a biphenyl linker.

Linker physical properties. The physical properties of linkers arenominally determined by the chemical constitution and bonding patternsof the linker, and linker physical properties impact the overallphysical properties of the candidate multibinding compounds in whichthey are included. A range of linker compositions is typically selectedto provide a range of physical properties (hydrophobicity,hydrophilicity, amphiphilicity, polarizability, acidity, and basicity)in the candidate multibinding compounds. The particular choice of linkerphysical properties is made within the context of the physicalproperties of the ligands they join and preferably the goal is togenerate molecules with favorable PK/ADME properties. For example,linkers can be selected to avoid those that are too hydrophilic or toohydrophobic to be readily absorbed and/or distributed in vivo.

Linker chemical functional groups. Linker chemical functional groups areselected to be compatible with the chemistry chosen to connect linkersto the ligands and to impart the range of physical properties sufficientto span initial examination of this parameter.

Combinatorial synthesis

Having chosen a set of n ligands (n being determined by the sum of thenumber of different attachment points for each ligand chosen) and mlinkers by the process outlined above, a library of (n!)m candidatedivalent multibinding compounds is prepared which spans the relevantmultibinding design parameters for a particular target. For example, anarray generated from two ligands, one which has two attachment points(A1, A2) and one which has three attachment points (B1, B2, B3) joinedin all possible combinations provide for at least 15 possiblecombinations of multibinding compounds:

A1- A1-A2 A1-B1 A1-B2 A1-B3 A2-A2 A2-B1 A2-B2 A1 A2- B1-B1 B1-B2 B1-B3B2-B2 B2-B3 B3-B3 B3

When each of these combinations is joined by 10 different linkers, alibrary of 150 candidate multibinding compounds results.

Given the combinatorial nature of the library, common chemistries arepreferably used to join the reactive functionalies on the ligands withcomplementary reactive functionalities on the linkers. The librarytherefore lends itself to efficient parallel synthetic methods. Thecombinatorial library can employ solid phase chemistries well known inthe art wherein the ligand and/or linker is attached to a solid support.Alternatively and preferably, the combinatorial libary is prepared inthe solution phase. After synthesis, candidate multibinding compoundsare optionally purified before assaying for activity by, for example,chromatographic methods (e.g., HPLC).

Analysis of array by biochemical analytical, pharmacological, andcomputational methods

Various methods are used to characterize the properties and activitiesof the candidate multibinding compounds in the library to determinewhich compounds possess multibinding properties. Physical constants suchas solubility under various solvent conditions and logD/clogD values aredetermined. A combination of NMR spectroscopy and computational methodsis used to determine low-energy conformations of the candidatemultibinding compounds in fluid media. The ability of the members of thelibrary to bind to the desired target and other targets is determined byvarious standard methods, which include radioligand displacement assaysfor receptor and ion channel targets, and kinetic inhibition analysisfor many enzyme targets. In vitro efficacy, such as for receptoragonists and antagonists, ion channel blockers, and antimicrobialactivity, are also determined. Pharmacological data, including oralabsorption, everted gut penetration, other pharmacokinetic parametersand efficacy data are determined in appropriate models. In this way, keystructure-activity relationships are obtained for multibinding designparameters which are then used to direct future work.

The members of the library which exhibit multibinding properties, asdefined herein, can be readily determined by conventional methods. Firstthose members which exhibit multibinding properties are identified byconventional methods as described above including conventional assays(both in vitro and in vivo.

Second, ascertaining the structure of those compounds which exhibitmultibinding properties can be accomplished via art recognizedprocedures. For example, each member of the library can be encrypted ortagged with appropriate information allowing determination of thestructure of relevant members at a later time. See: for example, Dower,et al., International Patent Application Publication No. WO 93/06121;Brenner, et al. , Proc. Natl. Acad. Sci., USA, 89:5181 (1992); Gallop,et al., U.S. Pat. No. 5,846,839; each of which are incorporated hereinby reference in its entirety. Alternatively, the structure of relevantmultivalent compounds can also be determined from soluble and untaggedlibaries of candidate multivalent compounds by methods known in the artsuch as those described by Hindsgaul, et al., Canadian PatentApplication No. 2,240,325 which was published on Jul. 11, 1998. Suchmethods couple frontal affinity chromatography with mass spectroscopy todetermine both the structure and relative binding affinities ofcandidate multibinding compounds to receptors.

The process set forth above for dimeric candidate multibinding compoundscan, of course, be extended to trimeric candidate compounds and higheranalogs thereof.

Follow-up synthesis and analysis of additional array(s)

Based on the information obtained through analysis of the initiallibrary, an optional component of the process is to ascertain one ormore promising multibinding “lead” compounds as defined by particularrelative ligand orientations, linker lengths, linker geometries, etc.Additional libraries are then generated around these leads to providefor further information regarding structure to activity relationships,These arrays typically bear more focused variations in linker structureto further optimize target affinity and/or activity at the target(antagonism, partial agoinism, etc.), and/or alter physical properties.By iterative redesign/analysis using the novel principles ofmultibinding design along with classical medicinal chemistry,biochemistry, and pharmacrology approaches, one is able to prepare andidentify optimal multibinding compounds that exhibit biologicaladvantage towards their targets and as therapeutic agents.

To further elaborate upon this procedure, suitable divalent linkersinclude, by way of example only, those derived from dicarboxylic acids,disulfonylhalides, dialdehydes, diketones, dihalides, diisocyanates,diamines, diols, mixtures of carboxylic acids, sulfonylhalides,aldehydes, ketones, halides, isocyanates, amines and diols. In eachcase, the carboxylic acid, sulfonylhalide, aldehyde, ketone, halide,isocyanate, amine and diol functional group is reacted with acomplementary functionality on the ligand to form a covalent linkage.Such complementary functionality is well known in the art as illustratedin the following table:

COMPLEMENTARY BINDING CHEMISTRIES First Reactive Group Second ReactiveGroup Linkage hydroxyl isocyanate urethane amine epoxide β-aminohydroxysulfonyl halide amine sulfonamide carboxyl acid amine amide hydroxylalkyl/aryl halide ether aldehyde amine/NaCNBH₃ amine ketoneamine/NaCNBH₃ amine amine isocyanate urea

The following table illustrates, by way of example, starting materials(identified as X-1 through X-418) that can be used to prepare linkersincorporated in the multibinding compounds of this invention utilizingthe chemistry described above. For example, 1,10-decanedicarboxylicacid, X1, can be reacted with 2 equivalents of a ligand carrying anamino group in the presence of a coupling reagent such as DCC to providea bivalent multibinding compound of formula (I) wherein the ligands arelinked via a 1,10-decanediamido linking group.

Representative ligands for use in this invention include, by way ofexample, those described above.

For example, L-1 can be an anti-seizure compound (e.g., lamotrigine,compounds 36 of Scheme J (described herein), carbamazepine and 4030W92);

L-2 can be a local anesthetic (e.g., lidocaine, and QX-314); and

L3 can be an anti-arrhythmic compound (e.g., mexilitene, tocainide, andflecainide).

Combinations of ligands (L) and linkers (X) per this invention include,by way example only, homo- and hetero-dimers wherein a first ligand isselected from L-1 through L-3 above and the second ligand and linker isselected from the following:

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L-3/X-114-L-3/X-115- L-3/X-116- L-3/X-117- L-3/X-118- L-3/X-119- L-3/X-120-L-3/X-121- L-3/X-122- L-3/X-123- L-3/X-124- L-3/X-125- L-3/X-126-L-3/X-127- L-3/X-128- L-3/X-129- L-3/X-130- L-3/X-131- L-3/X-132-L-3/X-133- L-3/X-134- L-3/X-135- L-3/X-136- L-3/X-137- L-3/X-138-L-3/X-139- L-3/X-140- L-3/X-141- L-3/X-142- L-3/X-143- L-3/X-144-L-3/X-145- L-3/X-146- L-3/X-147- L-3/X-148- L-3/X-149- L-3/X-150-L-3/X-151- L-3/X-152- L-3/X-153- L-3/X-154- L-3/X-155- L-3/X-156-L-3/X-157- L-3/X-158- L-3/X-159- L-3/X-160- L-3/X-161- L-3/X-162-L-3/X-163- L-3/X-164- L-3/X-165- L-3/X-166- L-3/X-167- L-3/X-168-L-3/X-169- L-3/X-170- L-3/X-171- L-3/X-172- L-3/X-173- L-3/X-174-L-3/X-175- L-3/X-176- L-3/X-177- L-3/X-178- L-3/X-179- L-3/X-180-L-3/X-181- L-3/X-182- L-3/X-183- L-3/X-184- L-3/X-185- L-3/X-186-L-3/X-187- L-3/X-188- L-3/X-189- L-3/X-190- L-3/X-191- L-3/X-192-L-3/X-193- L-3/X-194- L-3/X-195- L-3/X-196- L-3/X-197- L-3/X-198-L-3/X-199- L-3/X-200- L-3/X-201- L-3/X-202- L-3/X-203- L-3/X-204-L-3/X-205- L-3/X-206- L-3/X-207- L-3/X-208- L-3/X-209- L-3/X-210-L-3/X-211- L-3/X-212- L-3/X-213- L-3/X-214- L-3/X-215- L-3/X-216-L-3/X-217- L-3/X-218- L-3/X-219- L-3/X-220- L-3/X-221- L-3/X-222-L-3/X-223- L-3/X-224 L-3/X-225- L-3/X-226- L-3/X-227- L-3/X-228-L-3/X-229- L-3/X-230- L-3/X-231- L-3/X-232- L-3/X-233- L-3/X-234-L-3/X-235- L-3/X-236- L-3/X-237- L-3/X-238- L-3/X-239- L-3/X-240-L-3/X-241- L-3/X-242- L-3/X-243- L-3/X-244- L-3/X-245- L-3/X-246-L-3/X-247- L-3/X-248- L-3/X-249- L-3/X-250- L-3/X-251- L-3/X-252-L-3/X-253- L-3/X-254- L-3/X-255- L-3/X-256- L-3/X-257- L-3/X-258-L-3/X-259- L-3/X-260- L-3/X-261- L-3/X-262- L-3/X-263- L-3/X-264-L-3/X-265- L-3/X-266- L-3/X-267- L-3/X-268- L-3/X-269- L-3/X-270-L-3/X-271- L-3/X-272- L-3/X-273- L-3/X-274- L-3/X-275- L-3/X-276-L-3/X-277- L-3/X-278- L-3/X-279- L-3/X-280- L-3/X-281- L-3/X-282-L-3/X-283- L-3/X-284- L-3/X-285- L-3/X-286- L-3/X-287- L-3/X-288-L-3/X-289- L-3/X-290- L-3/X-291- L-3/X-292- L-3/X-293- L-3/X-294-L-3/X-295- L-3/X-296- L-3/X-297- L-3/X-298- L-3/X-299- L-3/X-300-L-3/X-301- L-3/X-302- L-3/X-303- L-3/X-304- L-3/X-305- L-3/X-306-L-3/X-307- L-3/X-308- L-3/X-309- L-3/X-310- L-3/X-311- L-3/X-312-L-3/X-313- L-3/X-314- L-3/X-315- L-3/X-316- L-3/X-317- L-3/X-318-L-3/X-319- L-3/X-320- L-3/X-321- L-3/X-322- L-3/X-323- L-3/X-324-L-3/X-325- L-3/X-326- L-3/X-327- L-3/X-328- L-3/X-329- L-3/X-330-L-3/X-331- L-3/X-332- L-3/X-333- L-3/X-334- L-3/X-335- L-3/X-336-L-3/X-337- L-3/X-338- L-3/X-339- L-3/X-340- L-3/X-341- L-3/X-342-L-3/X-343- L-3/X-344- L-3/X-345- L-3/X-346- L-3/X-347- L-3/X-348-L-3/X-349- L-3/X-350- L-3/X-351- L-3/X-352- L-3/X-353- L-3/X-354-L-3/X-355- L-3/X-356- L-3/X-357- L-3/X-358- L-3/X-359- L-3/X-360-L-3/X-361- L-3/X-362- L-3/X-363- L-3/X-364- L-3/X-365- L-3/X-366-L-3/X-367- L-3/X-368- L-3/X-369- L-3/X-370- L-3/X-371- L-3/X-372-L-3/X-373- L-3/X-374- L-3/X-375- L-3/X-376- L-3/X-377- L-3/X-378-L-3/X-379- L-3/X-380- L-3/X-381- L-3/X-382- L-3/X-383- L-3/X-384-L-3/X-385- L-3/X-386- L-3/X-387- L-3/X-388- L-3/X-389- L-3/X-390-L-3/X-391- L-3/X-392- L-3/X-393- L-3/X-394- L-3/X-395- L-3/X-396-L-3/X-397- L-3/X-398- L-3/X-399- L-3/X-400- L-3/X-401- L-3/X-402-L-3/X-403- L-3/X-404- L-3/X-405- L-3/X-406- L-3/X-407- L-3/X-408-L-3/X-409- L-3/X-410- L-3/X-411- L-3/X-412- L-3/X-413- L-3/X-414-L-3/X-415- L-3/X-416- L-3/X-417- L-3/X-418- and so on

METHODS OF PREPARATION

Linkers

The linker or linkers, when covalently attached to multiple copies ofthe ligands, provides a biocompatible, substantially non-immunogenicmultibinding compound. The biological activity of the multibinding Na⁺channel compound is highly sensitive to the geometry, composition, size,length, flexibility or rigidity, the presence or absence of anionic orcationic charge, the relative hydrophobicity/hydrophilicity, and similarproperties of the linker. Accordingly, the linker is preferably chosento maximize the biological activity of the compound. The linker may bebiologically “neutral,” i.e., not itself contribute any additionalbiological activity to the multibinding compound, or it may be chosen tofurther enhance the biological activity of the compound. In general, thelinker may be chosen from any organic molecule construct that orientstwo or more ligands for binding to the receptors to permit multivalency.In this regard, the linker can be considered as a “framework” on whichthe ligands are arranged in order to bring about the desiredligand-orienting result, and thus produce a multibinding compound.

For example, different orientations of ligands can be achieved byvarying the geometry of the framework (linker) by use of mono- orpolycyclic groups, such as aryl and/or heteroaryl groups, or structuresincorporating one or more carbon—carbon multiple bonds (alkenyl,alkenylene, alkynyl or alkynylene groups). The optimal geometry andcomposition of frameworks (linkers) used in the multibinding compoundsof this invention are based upon the properties of their intendedreceptors. For example, it is preferred to use rigid cyclic groups(e.g., aryl, heteroaryl), or non-rigid cyclic groups (e.g., cycloalkylor crown groups) to reduce conformational entropy when such may benecessary to achieve energetically coupled binding.

Different hydrophobic/hydrophilic characteristics of the linker as wellas the presence or absence of charged moieties can readily be controlledby the skilled artisan. For example, the hydrophobic nature of a linkerderived from hexamethylene diamine (H₂N(CH₂)₆NH₂) or related polyaminescan be modified to be substantially more hydrophilic by replacing thealkylene group with a poly(oxyalkylene) group such as found in thecommercially available “Jeffamines” (class of surfactants).

Different frameworks can be designed to provide preferred orientationsof the ligands. The identification of an appropriate framework geometryfor ligand domain presentation is an important first step in theconstruction of a multi binding agent with enhanced activity. Systematicspatial searching strategies can be used to aid in the identification ofpreferred frameworks through an iterative process. FIGS. 3A and 3Billustrate a useful strategy for determining an optimal frameworkdisplay orientation for ligand domains and can be used for preparing thebivalent compounds of this invention. Various alternative strategiesknown to those skilled in the art of molecular design can be substitutedfor the one described here.

As shown in FIGS. 3A and 3B, the ligands (shown as filled circles) areattached to a central core structure such as phenyldiacetylene (Panel A)or cyclohexane dicarboxylic acid (Panel B). The ligands are spaced apartfrom the core by an attaching moiety of variable lengths m and n. If theligand possesses multiple attachment sites (see discussion below), theorientation of the ligand on the attaching moiety may be varied as well.The positions of the display vectors around the central core structuresare varied, thereby generating a collection of compounds. Assay of eachof the individual compounds of a collection generated as described willlead to a subset of compounds with the desired enhanced activities(e.g., potency, selectivity). The analysis of this subset using atechnique such as Ensemble Molecular Dynamics will suggest a frameworkorientation that favors the properties desired.

The process may require the use of multiple copies of the same centralcore structure or combinations of different types of display cores. Itis to be noted that core structures other than those shown here can beused for determining the optimal framework display orientation of theligands. The above-described technique can be extended to trivalentcompounds and compounds of higher-order valency.

A wide variety of linkers is commercially available (e.g., Chem SourcesUSA and Chem Sources International; the ACD electronic database; andChemical Abstracts). Many of the linkers that are suitable for use inthis invention fall into this category. Others can be readilysynthesized by methods known in the art, and as described below.Examples of linkers include aliphatic moieties, aromatic moieties,steroidal moieties, peptides, and the like. Specific examples arepeptides or polyamides, hydrocarbons, aromatics, heterocyclics, ethers,lipids, cationic or anionic groups, or a combination thereof.

Examples are given below and in FIG. 4, but it should be understood thatvarious changes may be made and equivalents may be substituted withoutdeparting from the true spirit and scope of the invention. For example,properties of the linker can be modified by the addition or insertion ofancillary groups into the linker, for example, to change the solubilityof the multibinding compound (in water, fats, lipids, biological fluids,etc.), hydrophobicity, hydrophilicity, linker flexibility, antigenicity,stability, and the like. For example, the introduction of one or morepoly(ethylene glycol) (PEG) groups onto the linker enhances thehydrophilicity and water solubility of the multibinding compound,increases both molecular weight and molecular size and, depending on thenature of the unPEGylated linker, may increase the in vivo retentiontime. Further, PEG may decrease antigenicity and potentially enhancesthe overall rigidity of the linker.

Ancillary groups that enhance the water solubility/hydrophilicity of thelinker, and accordingly, the resulting multibinding compounds, areuseful in practicing this invention. Thus, it is within the scope of thepresent invention to use ancillary groups such as, for example, smallrepeating units of ethylene glycols, alcohols, polyols, (e.g., glycerin,glycerol propoxylate, saccharides, including mono-, oligosaccharides,etc.) carboxylates (e.g., small repeating units of glutamic acid,acrylic acid, etc.), amines (e.g., tetraethylenepentamine), and the liketo enhance the water solubility and/or hydrophilicity of themultibinding compounds of this invention. In preferred embodiments, theancillary group used to improve water solubility/hydrophilicity will bea polyether. In particularly preferred embodiments, the ancillary groupwill contain a small number of repeating ethylene oxide (—CH₂CH₂O—)units.

The incorporation of lipophilic ancillary groups within the structure ofthe linker to enhance the lipophilicity and/or hydrophobicity of thecompounds of Formula I is also within the scope of this invention.Lipophilic groups useful with the linkers of this invention include, butare not limited to, lower alkyl, aromatic groups and polycyclic aromaticgroups. The aromatic groups may be either unsubstituted or substitutedwith other groups, but are at least substituted with a group whichallows their covalent attachment to the linker. As used herein the term“aromatic groups” incorporates both aromatic hydrocarbons andheterocyclic aromatics. Other lipophilic groups useful with the linkersof this invention include fatty acid derivatives which may or may notform micelles in aqueous medium and other specific lipophilic groupswhich modulate interactions between the multibinding compound andbiological membranes.

Also within the scope of this invention is the use of ancillary groupswhich result in the compound of Formula I being incorporated into avesicle, such as a liposome, or a micelle. The term “lipid” refers toany fatty acid derivative that is capable of forming a bilayer ormicelle such that a hydrophobic portion of the lipid material orientstoward the bilayer while a hydrophilic portion orients toward theaqueous phase. Hydrophilic characteristics derive from the presence ofphosphato, carboxylic, sulfato, amino, sulfhydryl, nitro and other likegroups well known in the art. Hydrophobicity could be conferred by theinclusion of groups that include, but are not limited to long chainsaturated and unsaturated aliphatic hydrocarbon groups of up to 20carbon atoms and such groups substituted by one or more aryl,heteroaryl, cycloalkyl, and/or heterocyclic group(s). Preferred lipidsare phosphoglycerides and sphingolipids, representative examples ofwhich include phosphatidylcholine, phosphatidylethanolamine,phosphatidylserine, phosphatidylinositol, phosphatidic acid,pannitoyleoyl phosphatidylcholine, lysophosphatidylcholine,lysophosphatidyl-ethanolane, dipalimtoylphosphatidylcholine,dioleoylphosphatidyl-chohne, distearoyl-phosphatidylcholine anddilinoleoylphosphatidylcholine. Other compounds lacking phosphorus, suchas sphingolipid and glycosphingolipid families, are also within thegroup designated as lipid. Additionally, the amphipathic lipidsdescribed above may be mixed with other lipids including triglyceridesand sterols.

The flexibility of the linker can be manipulated by the inclusion ofancillary groups which are bulky and/or rigid. The presence of bulky orrigid groups can hinder free rotation about bonds in the linker, orbonds between the linker and the ancillary group(s), or bonds betweenthe linker and the functional groups. Rigid groups can include, forexample, those groups whose conformational freedom is restrained by thepresence of rings and/or π-bonds, for example, aryl, heteroaryl andheterocyclic groups. Other groups which can impart rigidity includepolypeptide groups such as oligo- or polyproline chains.

Rigidity can also be imparted electrostatically. Thus, if the ancillarygroups are either positively or negatively charged, the similarlycharged ancillary groups will force the linker into a configurationaffording the maximum distance between each of the like charges. Theenergetic cost of bringing the like charged groups closer to each other,which is inversely related to the square of the distance between thegroups, will tend to hold the linker in a configuration that maintainsthe separation between the like-charged ancillary groups. Further,ancillary groups bearing opposite charges will tend to be attracted totheir oppositely charged counterparts and potentially may enter intoboth inter- and intramolecular ionic bonds. This non-covalent mechanismwill tend to hold the linker in a conformation which allows bondingbetween the oppositely charged groups. The addition of ancillary groupswhich are charged, or alternatively, protected groups that bear a latentcharge which is unmasked, following addition to the linker, bydeprotection, a change in pH, oxidation, reduction or other mechanismsknown to those skilled in the art, is within the scope of thisinvention.

Bulky groups can include, for example, large atoms, ions (e.g., iodine,sulfur, metal ions, etc.) or groups containing large atoms, polycyclicgroups, including aromatic groups, non-aromatic groups and structuresincorporating one or more carbon—carbon π-bonds (i.e., alkenes andalkynes). Bully groups can also include oligomers and polymers which arebranched- or straight-chain species. Species that are branched areexpected to increase the rigidity of the structure more per unitmolecular weight gain than are straight-chain species.

In preferred embodiments, rigidity (entropic control) is imparted by thepresence of alicyclic (e.g., cycloalkyl), aromatic and heterocyclicgroups. In other preferred embodiments, this comprises one or moresix-membered rings. In still further preferred embodiments, the ring isan aryl group such as, for example, phenyl or naphthyl, or a macrocyclicring such as, for example, a crown compound.

In view of the above, it is apparent that the appropriate selection of alinker group providing suitable orientation, entropy andphysico-chemical properties is well within the skill of the art.

Eliminating or reducing antigenicity of the multibinding compoundsdescribed herein is also within the scope of this invention. In certaincases, the antigenicity of a multibinding compound may be eliminated orreduced by use of groups such as, for example, poly(ethylene glycol).

The Compounds of Formula I

As explained above, the multibinding compounds described herein comprise2-10 ligands attached covalently to a linker that links the ligands in amanner that allows their multivalent binding to ligand binding sites ofNa⁺ channels. The linker spatially constrains these interactions tooccur within dimensions, defined by the linker. This and other factorsincreases the biologic and/or therapeutic effect of the multibindingcompound as compared to the same number of ligands used in monobindingform.

The compounds of this invention are preferably represented by theempirical formula (L)_(p)(X)_(q) where L, X, p and q are as definedabove. This is intended to include the several ways in which the ligandscan be linked together in order to achieve the objective ofmultivalency, and a more detailed explanation is provided below.

As noted previously, the linker may be considered as a framework towhich ligands are attached. Thus, it should be recognized that theligands can be attached at any suitable position on this framework, forexample, at the termini of a linear chain or at any intermediateposition thereof.

The simplest and most preferred multibinding compound is a bivalentcompound which can be represented as L—X—L, where L is a ligand and isthe same or different and X is the linker. A trivalent compound couldalso be represented in a linear fashion, i.e., as a sequence of repeatedunits L—X—L—X—L, in which L is a ligand and is the same or different ateach occurrence, as is X. However, a trivalent compound can alsocomprise three ligands attached to a central core, and thus berepresented as (L)₃X, where the linker X could include, for example, anaryl or cycloalkyl group. Tetravalent compounds can be represented in alinear array:

L—X—L—X—L—X—L,

or a branched array

i.e., a branched construct analogous to the isomers of butane (n-butyl,iso-butyl, sec-butyl, and t-butyl) or a tetrahedral array, e.g.

where X and L are as defined herein. Alternatively, it could berepresented as an alkyl, aryl or cycloalkyl derivative as describedabove with four (4) ligands attached to the core liner.

The same considerations apply to higher multibinding compounds of thisinvention containing from 5-10 ligands. However, for multibinding agentsattached to a central linker such as an aryl, cycloalkyl or heterocyclylgroup, or a crown compound, there is a self-evident constraint thatthere must be sufficient attachment sites on the linker to accommodatethe number of ligands present; for example, a benzene ring could notaccommodate more than 6 ligands, whereas a multi-ring linker (e.g.,biphenyl) could accommodate a larger number of ligands.

The above described compounds may alternatively be represented as cyclicchains of the form:

and variants thereof.

All of the above variations are intended to be within the scope of theinvention defined by the formula (L)_(p)(X)_(q), Examples of bivalentand higher-order valency compounds of this invention are provided inFIGS. 5A to 5D.

With the foregoing in mind, a preferred linker may be represented by thefollowing formula:

—X^(a)—Z—(Y^(a)—Z)_(m)—Y^(b)—Z—X^(a)—

in which:

m is an integer of from 0 to 20;

X^(a) at each separate occurrence is selected from the group consistingof —O—, —S—, —NR—, —C(O)—, —C(O)O—, —C(O)NR—, —C(S), —C(S)O—, —C(S)NR—or a covalent bond where R is as defined below;

Z is at each separate occurrence is selected from the group consistingof alkylene, substituted alkylene, cycloalkylene, substitutedcylcoalkylene, alkenylene, substituted alkenylene, alkynylene,substituted alkynylene, cycloalkenylene, substituted cycloalkenylene,arylene, heteroarylene, heterocyclene, or a covalent bond;

Y^(a) and Y^(b) at each separate occurrence are selected from the groupconsisting of:

 —S—S— or a covalent bond;

 in which:

n is 0, 1 or 2; and

R, R′ and R″ at each separate occurrence are selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl,substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryland heterocyclic.

Additionally, tile linker moiety can be optionally substituted at anyatom therein by one or more alkyl, substituted alkyl, cycloalkyl,substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl,substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryland heterocyclic group.

In anther embodiment of this invention, the linker (i.e., X, X′ or X″)has the formula.

wherein

each R^(a) is independently selected from the group consisting of acovalent bond, alkylene, substituted alkylene and arylene;

each R^(b) is independently selected from the group consisting ofhydrogen, alkyl and substituted alkyl; and

n′ is an integer ranging from 1 to about 20.

In view of the above description of the linker, it is understood thatthe term “linker” when used in combination with the term “multibindingcompound” includes both a covalently contiguous single linker (e.g.,L—X—L) and multiple covalently non-contiguous linkers (L—X—L—X—L) withinthe multibinding compound.

As was previously discussed, the linker or linkers can be attached todifferent positions on the ligand molecule to achieve differentorientations of the ligand domains and thereby facilitate multivalency.For example, the positions that are potentially available for linking arepresentative ligand are indicated by arrows in the structure shown inFIG. 6. Preferred positions of attachment suggested by known SAR areillustrated in the reaction schemes described herein.

Certain Na⁺ channel ligands may be chiral and exhibit stereoselectivity.The most active enantiomers are preferably used as ligands in themultibinding compounds of this invention. The chiral resolution ofenantiomers is accomplished by well known procedures that result in theformation of diastereomeric derivatives or salts, followed byconventional separation by chromatographic procedures or by fractionalcrystallization (see, e.g., Bossert, et al., Angew. Chem. Int. Ed.,20:762-769 (1981) and U.S. Pat. No. 5,571,827 and references citedtherein). Chiral ligands are also readily available via asymmetricsynthesis.

The ligands are covalently attached to the linker using conventionalchemical techniques. The reaction chemistries resulting in such linkageare well known in the art and involve the coupling of reactivefunctional groups present on the linker and ligand. In some cases, itmay be necessary to protect portions of the ligand that are not involvedin linking reactions.

Preferably, the reactive functional groups on the linker are selectedrelative to the functional groups on the ligand that are available forcoupling, or can be introduced onto the ligand for this purpose in someembodiments, the linker is coupled to ligand precursors, with thecompletion of ligand synthesis being carried out in a subsequent step.Where functional groups are lacking, they can be created by suitablechemistries that are described in standard organic chemistry texts suchas J. March, Advanced Organic Chemistry, 4^(th) Ed. (Wiley-Interscience,N.Y., 1992). Examples of the chemistry for connecting ligands by alinker are shown in FIG. 7 where R₁ and R₂ represent a ligand and/or thelinking group. One skilled in the art will appreciate that syntheticallyequivalent coupling reactions can be substituted for the reactionsillustrated herein.

The linker to which the ligands or ligand precursors are attachedcomprises a “core” molecule having two or more functional groups withreactivity that is complementary to that of the functional groups on theligand. FIG. 4 illustrates the diversity of “cores” that are useful forvarying the linker size, shape, length, orientation, rigidity,acidity/basicity, hydrophobicity/hydrophilicity, hydrogen bondingcharacteristics and number of ligands connected. This pictorialrepresentation is intended only to illustrate the invention, and not tolimit its scope to the structures shown. In the Figures and reactionschemes that follow, a solid circle is used to generically represent acore molecule. The solid circle is equivalent to a linker as definedabove after reaction.

Preparation of Multibinding Compounds of the Invention

The multibinding compounds of this invention can be prepared fromreadily available starting materials using the following general methodsand procedures. It will be appreciated that where typical or preferredprocess conditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Any suitable compound that binds to Na⁺cancels can be used as a ligandin this invention. Typically, a compound selected for use as a ligandwill have at lease one functional group, such as an amino, hydroxyl,thiol or carboxyl group and the like, which allows the compound to bereadily coupled to the linker.

In the examples below, the following abbreviations have the followingmeanings. If an abbreviation is not defined, it has its generallyaccepted meaning.

Å Angstroms cm centimeter DCC dicyclohexyl carbodiimide DIPEAN,N-diisopropylethylamine DMA N,N-dimethylacetamide DMFN,N-dimethylformamide DMSO dimethylsulfoxide DPPAdiphenylphosphorylazide EDTA ethylenediaminetetraacetic acid g gram HPLChigh performance liquid chromatography MEM minimal essential medium mgmilligram MIC minimum inhibitory concentration min minute mL millilitermm millimeter mmol millimol N normal TEA triethylamine THFtetrahydrofuran μL microliters μm microns

The preferred co pounds of Formula I are bivalent. It should be noted,however, that the same techniques can be used to generate higher ordermultibinding compounds, i.e., the compounds of the invention where p is3-10.

Reactions performed under standard amide coupling conditions are carriedout in an inert polar solvent (e.g., DMF, DMA) in the presence of ahindered base (e.g., TEA, DIPEA) and standard amide coupling reagents(e.g., DPPA, PyBOP, HATU, DCC).

The following describes several methods for preparing multibindingcompounds employing ligands having the following structures A and B:

Structure A

wherein, Y is N or C—R¹; Z is N or C—R²; R³ and R⁴ are eachindependently amino, substituted amino, halogen, hydroxyl, ether,thioether, alkyl, R⁵,R⁶,R⁷,R⁸, and R⁹ are each independently hydrogen,halogen, amino, substituted amino, hydroxyl, ether, thioether,fluoroalkyl, allyl, W is preferably 2,3-dichlorophenyl (whereinR⁵═R¹═Cl, and R⁷═R⁸═R⁹═H) or 2,3,5-trichlorophenyl (wherein R⁵═R⁶═R⁸═Cl,and R⁷═R⁹═H).

For structure A, three preferred subclasses of compounds are thepyrimidine series, the triazine series, and the pyrazine series asdescribed as follows.

Subclass Structure Illustrative Examples Pyrimidine (Y = N, C—R¹, Z = N)

4030W92 (W = 2,3-dichlorophenyl (R¹ = CH₂F, and R³ = R⁴ = NH₂)

Sipatrigine (W = 2,3,5-trichlorophenyl, (R¹ = H, R⁴ = NH₂, and R³ =4-methyl- piperazin-1-yl) Triazine (Y = N, Z = N)

Lamotrigine (W = 2,3-dichlorophenyl, and R³ = R⁴ = NH₂) PyrazineGW273293 (Y = N, Z = C—R²) (W = 2,3,5-trichlorophenyl, and R³ = R⁴ =NH₂)

Structure B

wherein R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are independently hydrogen,alkyl, halogen, ether, thioether, carboyl derivatives, hydroxyl,fluoroalkyl, amino, substituted amino, Y is —(CH₂)_(n)— where n is aninteger from 1-4, O, S, NR (where R=H or alkyl), (—R¹⁰¹R¹⁰²)_(n)— wheren is an integer from 1-4 and R¹⁰¹ and R¹⁰² are independently hydrogen,lower alkyl, or substituted lower alkyl and Z is —(CH)_(n)—,((—R¹⁰¹R¹⁰²)— where n is an integer from 14 and R¹⁰¹ and R¹⁰² areindependently hydrogen, lower alkyl, or substituted lower alkyl.

For structure B, three preferred compounds are: (1) mexilitene: Y—O,Z—CH₂—CH(CH₃)—R¹²═R¹⁶═Me, R¹³═R¹⁴═H, R¹⁷═R¹⁸═H; (2) N-ethyl mexilitene:Y═O, Z═CH₂—CH(CH₃)—, R¹²═R¹⁶═Me, R¹³═R¹⁴═R¹⁵═H, R¹⁷═Et, R¹⁸═H; and (3) aphenoxymethyl piperidine deriviative as described in EP 869119 A1,wherein Y═O, Z═CH₂—CH(R¹⁹)—CH₂—, R¹²═R¹⁶═Me, R¹³═R¹⁵═H, R¹⁴═Br, R¹⁷═Meand R¹⁸ and R¹⁹, taken together, are —(CH)₃—.

It will be understood by those skilled in the art that the followingmethods may be used to prepare other multibinding compounds of thisinvention.

The strategies for preparing compounds of Formula I discussed aboveinvolve coupling the ligand directly to a homobifunctional core. Anotherstrategy that can be used with all ligands, and for the preparation ofboth bivalent and higher order multibinding compounds, is to introduce a‘spacer’ before coupling to a central core. Such a spacer can itself beselected from the same set as the possible core compounds.

Compounds of Formula I of higher order valency, i.e., p=3-10, can beprepared by simple extension of the above strategies. Specificallycompounds are prepared by coupling ligands to a central core bearingmultiple functional groups. The reaction conditions are the same asdescribed above for the preparation of bivalent compounds, withappropriate adjustments made in the molar quantities of ligand andreagents.

All of the synthetic strategies described above employ a step in whichthe ligand, attached to spacers or not, is symmetrically linked tofunctionally equivalent positions on a central core. Compounds ofFormula I can also be synthesized using an asymmetric linear approach.This strategy may be preferred when linking two or more ligands atdifferent points of connectivity or when preparing heterovalomers.

Representative syntheses of ligand precursors are illustrated in thereaction schemes and examples shown in FIG. 8 and described herein.

Scheme A illustrates the synthesis of a pyrimidine class compound. Asshown, compounds 10 and 11 first undergo a base-catalyzed ClaisenReaction followed by alkylation to produce compound 12 which in turn isreacted with compound 13 to yield the pyrimidine compound. Thistechnique for synthesizing monovalent compounds and for the synthesis ofcompound 12 are described, for example, PCT application WO97/09317,BP372934A2, EP372934B1.

Scheme B illustrates a synthesis of a bivalent pyrimidine compound ofthe Formula I that adapts the method shown in scheme A wherein a dimericguanidine compound (13b) is employed in place of the monomeric compound(13). Compounds of formula (13b) can be made by known techniques, forexample, by reacting diamine linker(5) with compound (15) (seeSynthesis, (6), 579-82; 1994). Alternatively, they may be synthesized byreacting diamines with cyanamide in water to yield compounds of Formula(13b), (See, for example, German Patent DE 4240981.) The preparation ofcompound (15) is described in Tetrahedron Lett., 34(21), 3389-92 (1993).By varying the substitution on the benzaldehyde precursor of W othercompounds of formula (12) can be synthesized.

Example 1 illustrates the preparation of bivalent compound 52 of FormulaI via scheme B. Specifically, to a solution of NaOEt (from 9.13 mmol ofsodium) in ethanol (20 mL) is added piperazinodiformamidinedihydrochloride (51) (8.22 mmol). After stirring for a further 10minutes, 2-(2,3,5-trichlorophenyl)-3-methoxyacrylonitrile (50) (19.2mmol) is added and the mixture is stirred at reflux for 4 hours. Themixture is left standing at room temperature overnight and thenfiltered. The filtrate is concentrated and the residue is purified bychromatography to afford the title product. Compound (51) is describedin CAS 17238-65-2.

Scheme C illustrates the general principle of using conventionalsynthetic techniques to introduce functional groups in the ligand whichcan then be interconverted into other functional groups or dimerized. Asshown, compounds 10 and 11a produce pyrimidine 14a via thebase-catalyzed Claisen Reaction and alkylation process of scheme A. Inthis case, R¹ contains an acetal which is hydrolyzed and reduced toalcohol (14b). This process is described in WO97/09317 for2,4-diamino-5-(2,3-dichlorophenyl)-6-hydroxymethyl pyrimidine. Others ofform (14b) can be made by varying substitution at W, R³, following thetechniques described in WO97/09317.

Scheme D illustrates the synthesis of a bivalent compound of Formula Iby direct dimerization of the alcohol (14b) by a process whereby thealcohol is coupled to dihalide linker (3).

Example 2 illustrates the preparation of (55), a compound of Formula Ivia scheme D. Specifically, a solution of 20 mmols of (53) in DMF with10 mmols of 1,4-dibromobutane (54) and 20 mmols of diisopropylethylamineis heated at 80° C. and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the desired product. The preparation of compound (53) is alsodescribed in WO97/09317.

Scheme E illustrates the synthesis of a bivalent compound of Formula Iby oxidizing of alcohol (14b) into the aldehyde (14c), followed bydimerization by reductive alkylation with diamine linker (5).

Example 3 illustrates the preparation of a compound of Formula I (58),via Scheme E. Specifically, alcohol (53) (100 mmol) is dissolved inCH₂Cl₂. Pyridinium chloroformate (110 mmol) is added in portions withstirring. The progress of the reaction is monitored by TLC. When judgedcomplete, the solution is filtered through a small plug of silica gel,then evaporated under vacuum. The residue is chromatographed to affordthe desired product (56).

Diamine (57) (2 mmol) is dissolved in THF (10 ml). Acetic acid (0.5 ml)is then added and the reaction is heated to reflux. Aldehyde (56) (1mmol) dissolved in THF (10 ml) is then added dropwise to the refluxingsolution over 60 minutes and the reaction is refluxed for a further 60minutes. At this point, NaBH(OAc)₃ is added in portions and the reactionis stirred at reflux for a further 2 hours. The reaction is allowed tocool and then is quenched with aqueous NH₄Cl solution until the pH ofthe solution is adjusted to pH 7.0 using either 1 M HCl or 1 M NaOH. Theproduct is extracted from this aqueous phase with EtOAc. The organiclayer is dried using Na₂SO₄, the drying agent is then filtered off andthe solvent removed in vacuo to provide the crude product. The desiredmaterial is purified from this mixture using reverse phase HPLC.

Scheme F illustrates the synthesis of a bivalent compound of the FormulaI by conversion of aldehyde 14c to the amine (14d), followed bydimerization via amide coupling to diacid linker (4).

Example 4 illustrates the preparation of a compound of Formula I (61)via Scheme F. Specifically, aldehyde (56) (1 mmol) dissolved in CH₂Cl₂(10 ml) is then added dropwise over 60 minutes to a refluxing solutionof ammonium acetate (3 mmol) and acetic acid and the reaction isrefluxed for a further 60 minutes. At this point, NaBH(OAc)₃ is added inportions and the reaction is stirred at relux for a further 2 hours. Thereaction is allowed to cool and then is quenched with aqueous NH₄Clsolution until the pH of the solution is adjusted to pH 7.0 using either1 M HCl or 1 M NaOH. The product is extracted from this aqueous phasewith EtOAc. The organic layer is dried using Na₂SO₄, the drying agent isthen filtered off and the solvent removed in vacuo to provide the crudeproduct. The desired material is purified from this mixture usingreverse phase HPLC.

A solution of (59) (2 mmols) and isophthalic acid (60) (1 mmol) inmethylene chloride is prepared under argon in a flask equipped withmagnetic stirrer and drying tube. To this solution is addeddicyclohexylcarbodiimide (solid, 2.1 mmols) while stirring at roomtemperature. The course of the reaction is followed by thin layerchromatography. When reaction has occurred, the reaction solution isdiluted with ethyl acetate and washed with water and with aqueousNa₂CO₃. The organic layer is dried (NaSO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compoundis obtained by purification of the crude product by use of HPLC.

Scheme G illustrates the synthesis of bivalent compounds of Formula Ifrom monovalent compounds in the pyrimidine class. (The references citedwith respect to schemes J and K for the pyrazines are applicable forschemes G, H and I.) As shown, reaction of compound 21 and 20 via aPd-catalyzed aryl coupling reaction yields monovalent compound 22 whichis then coupled to diamine (5) to form the bivalent compound.

Example 5 illustrates the preparation of (69), a compound of Formula Ivia Scheme G. Specifically, a mixture of (65) (30 mmol) in THF andtetrakis(triphenylphosphine)palladium(0) is stirred under nitrogen atroom temperature for 10 minutes. 2M aqueous sodium carbonate is added tothe mixture followed by a solution of 2,3-dichlorobenzene boronic acid(66) (30 mmol) in absolute ethanol and the mixture refluxed undernitrogen for 17 hours. A further equivalent of 2,3,5-trichlorobenzeneboronic acid in absolute ethanol is added and the mixture refluxed foran additional 7.50 hours. Finally, another equivalent of2,3,5-trichlorobenzene boronic acid in absolute ethanol is added to themixture and continued refluxing for 17 hours. The cooled mixture isevaporated in vacuo. The residue is dissolved in chloroform, washed withaqueous saturated sodium bicarbonate and water, dried over anhydrousmagnesium sulfate, filtered and the filtrate evaporated down in vacuo.The residue is purified by flash chromatography usingchloroform/methanol as the eluant to afford the desired product (67).

A solution of 72 mmols of (67) in DMF with 36 mmols of1,3-diaminopropane (68) and 20 mmols of diisopropylethylamine is heatedas necessary in a sealed vessel and the reaction followed by TLC. Whenjudged complete, the mixture is partitioned between ethyl acetate andwater and the organic phase washed with water, dried over sodium sulfateand the solvent removed in vacuo. The residue is purified bychromatography to afford the desired product. Compound (65) is describedin CAS 3993-80-4 and compound (66) is described in WO 98/38174.

Scheme H illustrates the synthesis of another bivalent compounds ofFormula I from monovalent compounds in the pyrimidine class. As shown,reaction of compound 23 and 20 via a Pd-catalyzed aryl coupling reactionyields monovalent compound 24 which is then coupled to diamine (5) toform the bivalent compound.

Example 6 illustrates the preparation of (73), a compound of Formula Ivia Scheme H. Specifically, a mixture of (70) (30 mmol) in THF andtetrakis(triphenylphosphine)palladium(0) is stirred under nitrogen atroom temperature for 10 minutes. 12M aqueous sodium carbonate is addedto the mixture followed by a solution of 2,3-dichlorobenzene boronicacid (66) (30 mmol) in absolute ethanol and the mixture refluxed undernitrogen for 17 hours. A further equivalent of 2,3,5-trichlorobenzeneboronic acid in absolute ethanol is added and the mixture refluxed foran additional 7.50 hours. Finally, another equivalent of2,3,5-trichlorobenzene boronic acid in absolute ethanol is added to themixture and continued refluxing for 17 hours. The cooled mixture isevaporated in vacuo. The residue is dissolved in chloroform, washed withaqueous saturated sodium bicarbonate and water, dried over anhydrousmagnesium sulfate, filtered and the filtrate evaporated down in vacuo.The residue is purified by flash chromatography usingchloroform/methanol as the eluant to afford the desired product (71).

A solution of 72 mmols of (71) in DMF with 36 mmols ofN,N′-dimethyl-1,3-propanediamine (72) and 20 mmols ofdiisopropylethylamine is heated as necessary in a sealed vessel and thereaction followed by TLC. When judged complete, the mixture ispartitioned between ethyl acetate and water and the organic phase washedwith water, dried over sodium sulfate and the solvent removed in vacuo.The residue is purified by chromatography to afford the desired product.Compound (70) is described in CAS 205672-25-9.

Scheme I illustrates the synthesis of another bivalent compound ofFormula I from monovalent compounds in the pyrimidine class. As shown,reaction of compound 25 and 20 via a Pd-catalyzed aryl coupling reactionyields monovalent compound 26 which is then coupled to diamine (5) toform the bivalent compound.

Example 7 illustrates the preparation of (78), a compound of Formula Ivia Scheme I. Specifically, a mixture of (74) (30 mmol) in THF andtetrakis(triphenylphosphine)palladium(0) is stirred under nitrogen atroom temperature for 10 minutes. 2M aqueous sodium carbonate is added tothe mixture followed by a solution of 2,3-dichlorobenzene boronic acid(66) (30 mmol) in absolute ethanol and the mixture refluxed undernitrogen for 17 hours. A further equivalent of 2,3,5-trichlorobenzeneboronic acid in absolute ethanol is added and the mixture refluxed foran additional 7.50 hours. Finally, another equivalent of2,3,5-trichlorobenzene boronic acid in absolute ethanol is added to themixture and continued refluxing for 17 hours. The cooled mixture isevaporated in vacuo. The residue is dissolved in chloroform, washed withaqueous saturated sodium bicarbonate and water, dried over anhydrousmagnesium sulfate, filtered and the filtrate evaporated down in vacuo.The residue is purified by flash chromatography usingchloroform/methanol as the eluant to afford the desired product (75).

A solution of 72 mmols of (75) in DMF with 36 mmols of piperazine (76)and 20 mmols of diisopropylethylamine is heated as necessary in a sealedvessel and the reaction followed by TLC. When judged complete, themixture is partitioned between ethyl acetate and water and the organicphase washed with water, dried over sodium sulfate and the solventremoved in vacuo. The residue is purified by chromatography to affordthe desired product (77).

A suspension of (77) (72 mmol) in absolute ethanol and ammonia (375 ml)is stirred and heated in an autoclave at 160° C. and 20 atm. for 16hours. The cooled mixture is evaporated in vacuo and extracted with hotmethanol. The combined methanol extracts are evaporated in vacuo. Theresidue is dissolved in hot chloroform, dried over anhydrous magnesiumsulfate, filtered and the filtrate evaporated in vacuo. The residue istriturated with 40-60° C. petroleum ether, filtered, and dried in vacuoto afford the desired product. Compound (74) is described in CAS13544-44-0. (Note that ins scheme I the coupling leaves two Cls (26);selective coupling at the more reactive position gives (27)).

Scheme J illustrates the synthesis bivalent compounds of Formula I frommonovalent compounds in the pyrazine class. As shown, reaction ofcompound 35 and 20 via a Pd-catalyzed aryl coupling reaction yieldsmonovalent compound 36 which is then coupled to diamine (5) to form thebivalent compound. This reaction is described in WO98/38174.

Example 8 illustrates the preparation of (82), a compound of Formula Ivia Scheme J. Specifically, a solution of 72 mmols of2-amino-6-chloro-3-(2,3,5-trichlorophenyl)pyrazine (80) in DMF with 36mmols of 1,3-diaminopropane (81) and 20 mmols of diisopropylethylamineis heated as necessary in a sealed vessel and the reaction followed byTLC. When judged complete, the mixture is partitioned between ethylacetate and water and the organic phase washed with water, dried oversodium sulfate and the solvent removed in vacuo. The residue is purifiedby chromatography to afford the desired product. Compound (80) isdescribed in WO 98/38174.

Scheme K illustrates the same process as shown in Scheme J adapted tocreate a compound (38) with the Cl in a different position in the ring.

Note for schemes G, H, I, J, and K the preferred compounds of Formula(20) (66) and (84) are described in WO98/38174, others are accessible byconventional synthesis (from an aromatic bromo compound).

Example 9 illustrates the preparation of (86), a compound of Formula Ivia Scheme K. Specifically, a mixture of2-chloro-3-bromo-6-acetamido-pyrazine (83) (30 mmol) in THF andtetrakis(triphenylphosphine)palladium(0) is stirred under nitrogen atroom temperature for 10 minutes. 2M aqueous sodium carbonate is added tothe mixture followed by a solution of 2,3,5-trichlorobenzene boronicacid (84) (30 mmol) in absolute ethanol and the mixture refluxed undernitrogen for 17 hours. A further equivalent of 2,3,5-trichlorobenzeneboronic acid in absolute ethanol is added and the mixture refluxed foran additional 7.50 hours. Finally, another equivalent of2,3,5-trichlorobenzene boronic acid in absolute ethanol is added to themixture and continued refluxing for 17 hours. The cooled mixture isevaporated in vacuo. The residue is dissolved in chloroform, washed withaqueous saturated sodium bicarbonate and water, dried over anhydrousmagnesium sulfate, filtered and the filtrate evaporated down in vacuo.The residue is purified by flash chromatography usingchloroform/methanol as the eluant to afford the desired product (85).

A solution of 72 mmols of (85) in DMF with 36 mmols of 1,4-diaminobutane(68) and 20 mmols of diisopropylethylamine is heated as necessary in asealed vessel and the reaction followed by TLC. When judged complete,the mixture is partitioned between ethyl acetate and water and theorganic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the desired product. Compound (83) is described in CAS173253-42-4.

Schemes L and M illustrate the general principle of using conventionalsynthetic techniques to introduce functional groups in the ligand whichcan then be interconverted into other functional groups or dimerized.

As shown, in scheme L illustrates the synthesis of a triazine classcompound. As shown, compounds 30 and 40 form triazine compound 41.

Scheme M illustrates the synthesis of a bivalent compound of the FormulaI from monovalent triazines that encompass lamotrigine. The synthesis oflamotrigine is further described in WO96/20934. As shown, followingsynthesis of thiol compound 41a from 30 and 40a, the thiol compound ismethylated to produce compound 42b. The product formed by oxidation ofcompound 42b is coupled to diamine linker (5) to produce the bivalentcompound.

EXAMPLE 10 Preparation of (91), a Compound of Formula I via Scheme M

A solution of 2,3,5-trichlorbenzoyl cyanide (87) (13 mmol) is dissolvedin acetonitrile and added dropwise to a suspension of (40a) (39 mmol) indilute sulphuric acid. The temperature is maintained below 30° C. Themixture is stirred at room temperature for 3 days. The solid isfiltered, washed with water and sucked dry. A suspension of the solid ina 10% solution of sodium hydroxide pellets in water is stirred at roomtemperature for 1 hour. The solid is filtered, washed with water anddried in vacuo. The solid is refluxed with hot n-propanol for 1.5 hours,filtered and dried in vacuo at 80° C. to afford the desired product(88).

A solution of 72 mmols of (88) in DMF with 72 mmols of methyl iodide and72 mmols of diisopropylethylamine is heated at 40° C. for 12 hours. Thereaction mixture is concentrated and chromatographed to afford thedesired product (89).

A solution of 60 mmols of (89) in dichloromethane with 120 mmols ofm-chloroperoxybenzoic acid is stirred at room temperature for 12 hours.The reaction mixture is concentrated and chromatographed to afford thedesired product. A solution of 50 mmols of the resulting product in DNwith 50 mmols diisopropylethylamine and 25 mmols of 4,4′-bipiperidinedihydrochloride (90) is heated at 120° C. for 12 hours in a sealedvessel. The reaction mixture is concentrated and chromatographed toafford the desired product. Compound (87) is reported in EP0459829(A1l).

Scheme N illustrates the synthesis of a bivalent compound of the FormulaI from monovalent triazine 42d which is produced by chlorination ofcompound 42c.

EXAMPLE 11 Preparation of (94) a Compound of Formula I via Scheme N

Alcohol (92) (5 mmol) is dissolved in CH₂Cl₂ at 0° C. and CBr₄ (12 mmol)is added. A solution of PPh₃ (15 mmol) in CH₂Cl₂ is added. The progressof the reaction is monitored by TLC. When judged complete, the solventis removed under vacuum and the residue is chromatographed to afford thedesired product (93).

A solution of 72 mmols of (93) in DMF with 36 mmols of1,3-cyclohexanebis(methylamine) and 20 mmols of diisopropylethylamine isheated as necessary in a sealed vessel and the reaction followed by TLC.When judged complete, the mixture is partitioned between ethyl acetateand water and the organic phase washed with water, dried over sodiumsulfate and the solvent removed in vacuo. The residue is purified bychromatography to afford the desired product. Compound (92) is describedin WO 96/20934.

Schemes O, P, and Q illustrate the general principle of linking throughthe ligand phenyl ring, with a functional group introduced in thisposition via several approaches. A different approach is shown for eachclass below.

Scheme O illustrates the synthesis of a compound of Formula I frommonovalent pyrimidine ligands that are coupled by dialdehyde linker (6).The pyrimidine is produced using a nitro-substituted starting material(10a) via the process of scheme A to yield nitro-substituted (14c).Aniline compound 14(f) is produced by reduction which is then dimerized.

EXAMPLE 12 Preparation of (98), a Compound of Formula I via Scheme O

A solution of (95) (0.0007M) in acetic acid (12 ml)/methanol (1 ml) isreduced under an atmosphere of hydrogen in the presence of PtO₂ (0.12g). The mixture is filtered and the filtrate is concentrated. Theresidue is neutralized with saturated NaHCO₃ solution and the product isextracted with ethylacetate, bulked, dried (MgSO₄) and evaporated toafford the desired product (96).

Compound (96) (2 mmol) is dissolved in THF (10 ml). Acetic acid (0.5 ml)is then added and the reaction is heated to reflux. Phthalaldehyde (1mmol) dissolved in THF (10 ml) is then added dropwise to the refluxingsolution over 60 minutes and the reaction is refluxed for a further 60minutes. At this point, NaBH(OAc)₃ (1 mmol) is added in portions and thereaction is stirred at relux for a further 2 hours. The reaction isallowed to cool and then is quenched with aqueous NH₄Cl solution untilthe pH of the solution is adjusted to pH 7.0 using either 1 M HCl or 1 MNaOH. The product is extracted from this aqueous phase with EtOAc. Theorganic layer is dried using NaSO₄, the drying agent is then filteredoff and the solvent removed in vacuo to provide the crude product. Thetitle compound is purified from this mixture using reverse phase HPLC.

Scheme P illustrates the synthesis of a compound of Formula I frommonovalent pyrazine ligands that are coupled by dihalide linker(3). Thepyrazine 47 is synthesized from a nitro-substituted starting material(45) via the process of Scheme J which initially yieldsnitro-substituted compound (46), which is then reduced and the anilineis used in dimerization.

EXAMPLE 13 Preparation of (134), a Compound of Formula I via Scheme P

A solution of (131) (0.0007M) in acetic acid (12 ml)/methanol (1 ml) isreduced under an atmosphere of hydrogen in the presence of PtO₂ (0.12g). The mixture is filtered and the filtrate is concentrated. Theresidue is neutralized with saturated NaHCO₃ solution and the product isextracted with ethylacetate, bulked, dried (MgSO₄) and evaporated toafford the desired product (132).

Compound (132) (1 mmol) is dissolved in DMF (5 mL) and treatedsequentially with 0.5 mmol α,α-bromo-o-xylene (133) and 2 mmol powderedpotassium carbonate. The mixture is heated as necessary to effectreaction, which is monitored by TLC. When judged complete, the mixtureis partitioned between ethyl acetate and water and the organic phasewashed with water, dried over, sodium sulfate and the solvent removed invacuo. The residue is purified by chromatography to afford the titlestructure.

Scheme Q illustrates the synthesis of a compound of Formula I frommonovalent triazine ligands that are coupled by dicarboxylic acid linker(4). The triazine 41c is formed by nitration of compound (41). Triazine41c is then reduced and the aniline 41d is used in dimerization.

EXAMPLE 14 Preparation of (101), a Compound of Formula I via Scheme Q

A solution of (99) (2 mmols) and adipic acid (100) (1 mmol) in methylenechloride is prepared under argon in a flask equipped with magneticstirrer and drying tube. To this solution is addeddicyclohexylcarbodiimide (solid, 2.1 mmols) while stirring at roomtemperature. The course of the reaction is followed by thin layerchromatography. When reaction has occurred, the reaction solution isdiluted with ethyl acetate and washed with water and with aqueousNa₂CO₃. The organic layer is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compoundis obtained by purification of the crude product by use of HPLC.

Schemes R, S, T, and U illustrate the synthesis of bivalent compounds ofFormula I from monovalent compounds of structure B. In each case, thelinkage is from the [N] group of a first ligand to the [N] group of asecond ligand.

Scheme R illustrates the coupling of a monovalent compound with dihalide3.

EXAMPLE 15 Preparation of (107), a Compound of Formula I via Scheme R

A solution of 20 mmols of (R)-N-ethyl mexiletine (105) in DMF with 10mmols of 1,6-dibromohexane (106) and 20 mmols of potassium carbonate isheated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the title structure. Compound (105) is reported in WO 97/27169.

EXAMPLE 16 Preparation of (109), a Compound of Formula I via Scheme R

A solution of 30 mmols of (R)-N-ethyl mexiletine (105) in DMF with 10mmols of 1,3,5-tri(bromoethyl)benzene (108) and 20 mmols of potassiumcarbonate is heated as necessary and the reaction followed by TLC. Whenjudged complete, the mixture is partitioned between ethyl acetate andwater and the organic phase washed with water, dried over sodium sulfateand the solvent removed in vacuo. The residue is purified bychromatography to afford the tide structure. Compound (105) is reportedin WO 97/27169 and compound (108) is described in CAS 18226-42-1.

EXAMPLE 17 Preparation of (114), a Compound of Formula I via Scheme R

A solution of 20 mmols of (R)-mexiletine (110) in DMF with 20 mmols of1,2-bis-(2-bromoethoxy)ethane (113) and 20 mmols of potassium carbonateis heated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the title structure. Compound (113) is described in CAS31255-10-4.

EXAMPLE 18 Preparation of (120), a Compound of Formula I via Scheme R

A solution of 20 mmols of(S)-3-(4-bromo-2,6-dimethylphenoxymethyl)piperdine (118) in DMF with 10mmols of 2-bromoethyl ether (119) and 20 mmols of potassium carbonate isheated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the title structure. Compound (118) is reported in EP 0869119 A1.

Scheme S illustrates the coupling of another monovalent compound withdihalide 3.

EXAMPLE 19 Preparation of (112), a Compound of Formula I via Scheme S

A solution of 20 mmols of (R)-mexiletine (110) in THF is treated with 20mmols of trifluoroacetic anhydride and 20 mmols of triethylamine. After1 hour, the solvent is removed in vacuo and the residue is partitionedbetween ethyl acetate and water. The organic layer is washed withadditional water, dried over anhydrous sodium sulfate, filtered and thesolvent removed under reduced pressure to afford the crudetrifluoroacetamide of (R)-mexiletine.

A solution of the above crude trifluoroacetamide in anhydrous THF iscooled under nitrogen to −78 C and treated dropwise with 20 mL of 1 NLDA in THF. The temperature is raised to −20 C and a solution of 10mmols of α′,α′-dibromo-p-xylene (111) is added and the reaction followedby TLC. When judged complete, 30 mL of 1 N NaOH is added and thetemperature is raised to 60 C until the trifluoroacetamide is removed asindicated by TLC. The reaction mixture is partitioned between ethylacetate and water. The organic layer is washed with additional water,dried over anhydrous sodium sulfate, filtered, and concentrated underreduced pressure. The residue is purified by chromatography to affordthe title structure.

Scheme T illustrates the coupling of two monovalent molecules with twodiamine linkers 5 to a linkage involving two linkers.

EXAMPLE 20 Preparation of (117), a Compound of Formula I via Scheme T

A solution of 20 mmols of 2-(2-bromopropoxy)-1,3-dimethylbenzene (115)in DMF with 10 mmols of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane(4,13-diaza-18-crown-6) (116) and 20 mmols of potassium carbonate isheated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography andseparation of the stereoisomers by known technique to afford the titlestructure. Compound (115) is described in CAS 96656-46-1.

The following Examples 21-24 (as illustrated in FIG. 8) describe thesyntheses of heterodimers comprising two nonidentical ligands.

EXAMPLE 21 Preparation of (122), a Compound of Formula I

A solution of 20 mmols of (R)-N-ethyl mexiletine (105) in DMF with 20mmols of 1,4-dibromobutane (54) and 20 mmols of potassium carbonate isheated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the desired product (121).

A solution of 20 mmols of(S)-3-(4-bromo-2,6-dimethylphenoxymethyl)piperdine (118) in DMF with 20mmols of the above product (121) and 20 mmols of potassium carbonate isheated as necessary and the reaction followed by TLC. When judgedcomplete, the mixture is partitioned between ethyl acetate and water andthe organic phase washed with water, dried over sodium sulfate and thesolvent removed in vacuo. The residue is purified by chromatography toafford the title structure. Compound (105) is reported in WO 97/27169.Compound (118) is reported in EP 0869119 A1.

EXAMPLE 22 Preparation of (123), a Compound of Formula I

A solution of 20 mmols of 2-(2-bromopropoxy)-1,3-dimethylbenzene (115)in DMF with 20 mmols of(S)-3-(4-bromo-2,6-dimethylphenoxymethyl)piperidine (118) and 20 mmolsof potassium carbonate is heated as necessary and the reaction followedby TLC. When judged complete, the mixture is partitioned between ethylacetate and water and the organic phase washed with water, dried oversodium sulfate and the solvent removed in vacuo. The residue is purifiedby chromatography to afford the title structure. Compound (115) asdescribed in CAS 96656-46-1 and compound (118) is reported in EP 0869119A1.

EXAMPLE 23 Preparation of (126), a Compound of Formula I

A solution of 10 mmols of (67) in 20 mL DMF is treated sequentially with15 mmols of diisopropylethylamine and 10 mmols of 1,4-diaminobutane(124). The solution is heated as necessary in a sealed vessel and thereaction followed by TLC. When it is judged complete, the mixturecontaining compound (125) is treated with 15 mmols additionaldiisopropylethylamine and 10 mmols of (80). The reaction is furtherheated as necessary and monitored by TLC until judged complete. Themixture is partitioned between ethyl acetate and water and the organicphase washed with water, dried over sodium sulfate and the solventremoved in vacuo. The residue is purified by chromatography to affordthe title compound.

Compound (80) is described in WO 98/38174.

EXAMPLE 24 Preparation of (127), a Compound of Formula I

A solution of 10 mmols of (118) in 20 mL DMF is treated sequentiallywith 30 mmols diisopropylethylamine and 20 mmols of (80). The solutionis heated as necessary in a sealed vessel and the reaction followed byTLC. When judged complete, the mixture is partitioned between ethylacetate and water and the organic phase washed with water, dried oversodium sulfate and the solvent removed in vacuo. The residue is purifiedby chromatography to afford the title compound. Compound (118) isreported in EP 0869119 A1 and compound (80) is described in WO 98/38174.

EXAMPLES 25-53

The syntheses of starting materials and multibinding compounds arefurther described in the following Examples 25 through 53.

In general, unless noted otherwise, starting material (includingdi-amines, di-aldehydes, di-halides, amines, alkyl halides, aldehydes,and etc.) and solvents were purchased from commercial suppliers(Aldrich, Fluka, Sigma, and etc.), and used without furtherpurification. Reactions were run under nitrogen atmosphere, unless notedotherwise such as in hydrogenation reaction. Progress of reactionmixtures was monitored by thin layer chromatography (TLC), analyticalhigh performance liquid chromatography (anal. HPLC), and massspectrometry, the details of which are given below and separately inspecific examples of reactions. Reaction mixtures were worked up asdescribed specifically in each reaction; Reaction mixtures were workedup as described specifically in each reaction; commonly it was purifiedby flash column chromatography with silica gel. Other purificationmethods include preparative TLC, temperature-, and solvent-dependentcrystallization, precipitation, and distillation. In addition, reactionmixtures were routinely purified by preparative HPLC: a general protocolis described below. Characterization of reaction products was routinelycarried out by mass and ¹H-NMR spectrometry. For NMR, samples weredissolved in deuterated solvent (CD₃OD, CDCl₃, or DMSO-d₆), and ¹H-NMRspectra were acquired with a Varian Gemini 2000 instrument (300 MHz)under standard observe parameters. Mass spectrometric identification ofcompounds was performed by an electrospray ionization method (ESMS) witha Perkin Elmer instrument (PE SCIEX API 150 EX).

A general protocol for analytical HPLC: Each crude compound wasdissolved in 50% MeCN/H₂O (with 0.1% TFA) at 0.5-1.0 mg/mLconcentration, and was analyzed by using anal. HPLC: 1) reversed-phaseanalytical column, Bonus-RP (2.1×50 mm; ID=5 μm); 2) flow rate: 0.5mL/min; 3) 10% MeCN/H₂O (0.1% TFA) (0-0.5 min), 10 to 70% (lineargradient; 0.5-5 min); 4) detection: 214, 254, and 280 nm.

A general protocol for preparative HPLC purification: Crude compoundswere dissolved in 50% MeCN/H₂O (with 0.1% TFA) at 30-45 mg/mLconcentration, filtered, and injected into a reversed-phase preparativecolumn. Following represents a typical example among variouspurification conditions: 1) column; YMC Pack-Pro C18 (50a×20 mm; ID=5μm); 2) linear gradient: 10 to 60% MeCN (0.1% TFA)/H₂O (0.1% TFA) over50 min; 3) flow rate: 40 mL/min; 4) detection: 214, 254, or 280 nm.

EXAMPLE 25 Synthesis of Mexiletine Dimers: Reductive Amination ofPrecursor Ketone 1 with bis-primary Amine via the Following Scheme

reagents and conditions: i) chloroacetone, K₂CO₃, KI, DMF, 80° C.; ii)H₂N-L-NH₂, EtOH rt; iii) NaBH₄, r.t.

To a stirred, suspension of 2,6-dimethylphenol (12.216 g, 100 mmol),potassium carbonate (14.0 g, 100 mmol), and potassium iodide (2.0 g,catalytic amount) in 400 mL of DMF at 80° C., was added dropwisely over30 min 12.0 mL (150 mmol) of chloroacetone. After completion of theaddition, the mixture was stirred and heated at 80° C. for 12 h. Aftercooling, the mixture was filtered, and concentrated in vacuo to give atarry residue. It was partitioned between ethyl acetate and water. Theorganic phase was dried over Na₂SO₄, and concentrated to afford the oilyresidue. Treatment with hexanes caused precipitation of brown solid,which was filtered off. The filtrate was passed through a pad of basicalumna to remove remaining phenol. Filtrates were concentrated toprovide the product ketone 1 as clear oil. ¹H-NMR (CD₃OD, 299.96 MHz): δ(ppm) 7.06-6.95 (d, 2H), 6.90-6.84 (dd, 1H), 4.49 (s, 2H), 2.23 (s, 9H).Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=3.56 min.

Additional dimer compounds (1-72) having the following structure A withdifferent linkers were synthesized using the above scheme.

The following Table A lists 72 linkers that can be employed. For linkers1-20 and 61-72, R of structure is H; for 21-46 R is a methyl group; andfor 47-60 R is an ethyl group. Each numbered compound refers to a dimerthat is linked by the linker in the table with the same number. Forexample, compound 1 would be linked by linker 1.

TABLE A No. Linker R 1 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— H 2 —(CH₂)₃—O—(CH₂)₃—H 3 —CH₂—Z—CH₂— where Z = 1,3-cyclohexyl H 4 —CH₂—CH(OH)—CH₂— H 5

H 6 —CH₂—Z—CH₂— where Z = 1,4-cyclohexyl H 7 —(CH₂)₅— H 8 —(CH₂)₆— H 9—(CH₂)₂—S—(CH₂)₂— H 10 —(CH₂)₃—O—(CH₂)₁₀—O—(CH₂)₃— H 11—(CH₂)₃—Z—(CH₂)₃— where Z = 2,4,8,10-tetraoxa- Hspiro[5.5]undecan-3,9-yl 12 —(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— H 13—(CH₂)₃—O—(CH₂)₄—O—(CH₂)₃— H 14 —(CH₂)₃—N(CH₃)—(CH₂)₃— H 15 —CH₂—Z—CH₂—where Z = trans-1,4-cyclohexyl H 16 —CH₂—CH(CH₃)—CH₂—C(CH₃)₂—(CH₂)₂— H17 —Z— where Z = 2,7-[9H-fluorene] H 18 —Z—C(CH₃)₂—Z—C(CH₃)₂—Z— where Z= 1,4-phenyl H 19 —Z—CH₂—Z— where Z = 1,4-phenyl H 20—(CH₂)₂—N(CH₃)—(CH₂)₂— H 21 —(CH₂)₉— Me 22 —CH₂—Z—CH₂— where Z =1,4-phenyl Me 23 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Me 24 —CH₂—CH═CH—CH₂— (transisomer) Me 25 —CH₂—Z—CH₂— where Z = (−)-trans-2,2-dimethyl- Me[1,3]dioxolan-4,5-yl 26 —(CH₂)₁₀— Me 27 —(CH₂)₁₁— Me 28 —(CH₂)₁₂— Me 29—(CH₂)₁₆— Me 30 —CH₂—CH(OH)—CH₂— Me 31 —CH₂—C═C—CH₂— Me 32 —(CH₂)₇— Me33 —(CH₂)₈— Me 34 —CH₂—Z—CH₂— where Z = 2,3-quinoxalinyl Me 35—CH₂—CH(CH₂OH)— Me 36 —CH₂—C(CH₂)—CH₂— Me 37 —Z— where Z =4,5-[1,3]dioxolan-2-one Me 38 —CH₂—Z—CH₂— where Z = 1,3-phenyl Me 39—(CH₂)₂—O—CH₂—O—(CH₂)₂— Me 40 —(CH₂)₄—O—(CH₂)₄— Me 41—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Me 42 —(CH₂)₂—O—C(O)—O—(CH₂)₂— Me 43—CH₂—CH[C(O)—O—CH₂CH₃]— Me 44 —CH₂—C(O)—O—(CH₂)₂—O—C(O)—CH₂— Me 45—(CH₂)₂—NH—C(O)—C(O)—NH—(CH₂)₂— Me 46 —(CH₂)₃— Me 47—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Et 48 —CH₂—Z—CH₂— where Z =(−)-trans-2,2-dimethyl- Et [1,3]dioxolan-4,5-yl 49 —(CH₂)₁₀— Et 50—(CH₂)₁₁— Et 51 —(CH₂)₁₂— Et 52 —(CH₂)₁₆— Et 53 —(CH₂)₇— Et 54—CH₂—Z—CH₂— where Z = 1,3-phenyl Et 55 —CH₂—Z—CH₂— where Z = 1,4-phenylEt 56 —(CH₂)₂—O—CH₂—O—(CH₂)₂— Et 57 —(CH₂)₄—O—(CH₂)₄— Et 58—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Et 59 —CH₂—C(O)—O—(CH₂)₂—O—C(O)—CH₂—Et 60 —(CH₂)₃— Et 61 —CH₂—Z—CH₂— where Z = 2,5-thiophenyl H 62—CH₂—Z—O—(CH₂)₂—O—Z—CH₂— where Z = 1,2- H phenyl 63—CH₂—Z—O—(CH₂)₆—O—Z—CH₂— where Z = 1,2- H phenyl 64—CH₂—Z—O—(CH₂)₃—O—Z—CH₂— where Z = 1,2- H phenyl 65 —CH₂—Z—CH₂— where Z= 2,3-thiophenyl H 66 —CH₂—Z—CH₂— where Z = 2,6-pyridinyl H 67—CH₂—Z—CH₂— where Z = 2-hydroxy-5-methyl-phenyl- H 1,3-yl 68—CH₂—Z—O—(CH₂)₂—O—Z—CH₂— where Z = 4- H methoxy-phenyl-1,3-yl 69—CH₂—Z—CH₂— where Z = 4-hydroxy-phenyl-1,3-yl H 70 —CH₂—Z—CH₂— where Z =2,2′-dihydroxy-3,3′- H dimethoxy-biphenyl-5,5′-yl 71 —CH₂—Z—O—Z—CH₂—where Z = 1,2-phenyl H 72 —CH₂—Z—CH₂— where Z = 1,3-phenyl H

General procedure for the synthesis of compound 1: To a solution ofcompound 1 (35.6 mg, 0.2 mmol) in 200 μL of anhydrous ethanol, was addeda solution of 1,8-diamino-3,6-dioxaoctane (14.8 mg, 0.1 mmol) in 200 μLof anhydrous ethanol. The mixture was shaken for 12 h at 25° C., andfollowed by addition of a solution of NaBH₄ (15.2 mg, 0.4 mmol) inethanol and shaking the mixture for 2 h at 25° C. The mixture was thenquenched with a solution of 5% trifluoroacetic acid in 50% aqueousacetonitrile, and concentrated under reduced pressure. The residue wasdissolved in 1 mL of a 1:1 mixture of acetonitrile and water (with 0.1%trifluoroacetic acid). The crude product was purified by preparativereversed phase HPLC. Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=4.14 min. ESMS (C₂₈H₄₄N₂O₄): calcd. 472.67; obsd. 473.4 [M+H]⁺.

Compound 2 was prepared in an analogous manner frombis-(3-aminopropyl)ether. Retention time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=4.61 min. ESMS (C₂₈H₄₄N₂O₃): calcd. 456.67; obsd. 457.4[M+H]⁺.

Compound 3 was prepared in an analogous manner from1,3-cyclohexane-bis-(methylamine). Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min) 4.60 min. ESMS (C₃₀H₄₆N₂O₂): calcd. 466. 71; obsd.467.4 [M+H]⁺.

Compound 4 was prepared in an analogous manner from1,3-diamino-2-propanol. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.12 min, 4.32 min. (mixture of diastereomers). ESMS(C₂₅H₃₈N₂O₃): calcd. 414.59; obsd. 415.4 [M+H]⁺.

Compound 5 was prepared in an analogous manner from1,4-bis-(3-aminopropyl)piperazine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=3.70 min. ESMS (C₂₅H₃₈N₂O₃): calcd. 524.79; obsd.525.8 [M+H]⁺.

Compound 6 was prepared in an analogous manner from1,4-cyclohexane-bis-methylamine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min) 4.30 min, 4.53 min (mixture of isomers). ESMS(C₃₀H₄₆N₂O₂): calcd. 466.71; obsd. 467.4 [M+H]⁺.

Compound 7 was prepared in an analogous manner from 1,5-diaminopentane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.18 min. 4.53min (mixture of isomers). ESMS (C₂₇H₄₂N₂O₂): calcd. 426.64; obsd. 427.4[M+H]⁺.

Compound 8 was prepared in an analogous manner from 1,6-diaminohexane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min) 4.26 min, 4.53min (mixture of isomers). ESMS (C₂₈H₄₄N₂O₂): calcd. 440.67; obsd. 441.4[M+H]⁺.

Compound 9 was prepared in an analogous manner from2,2′-thiobis(ethylamine). Retention time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=4.53 min. ESMS (C₂₆H₄₀N₂O₂S): calcd. 444.68; obsd. 441.4[M+H]⁺.

Compound 10 was prepared in an analogous manner from3,3′-(decamethylenedioxy)-bis-(propylamine). Retention time (anal. HPLC:10-70% MeCN/H₂O over 5 min)=4.98 min. ESMS (C₃₈H₆₄N₂O₄): calcd. 612.94;obsd. 613.6 [M+H]⁺.

Compound 11 was prepared in an analogous manner from3,9-bis(3-aminopropyl)-2,4,8,10-tetraoxapriro(5,5)undecane, Retentiontime (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.53 nm. ESMS (C₃₅HN₂O₆):calcd. 598.82; obsd. 599.4 [M+H]⁺.

Compound 12 was prepared in an analogous manner from4,7,10-trioxa-1,13-tridecanediamine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.17 min. ESMS (C₃₂H₂N₂O₅): calcd. 544.78; obsd.545.4 [M+H]⁺.

Compound 13 was prepared in an analogous manner from4,9-dioxa-1,12-dodecaneamine. Retention Time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.22 min, 4.52 min (mixture of isomers). ESMS(C₃₂H₅₂N₂O₄): calcd. 528.78; obsd. 529.4 [M+H]⁺.

Compound 14 was prepared in an analogous manner fromN,N-bis(3-aminopropyl)methylamine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=3.85 min. ESMS (C₂₉H₄₇N₃O): calcd. 469.71; obsd.470.2 [M+H]⁺.

Compound 15 was prepared in an analogous manner fromtrans-1,4-diaminocyclohexane. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=3.98 min. ESMS (C₂H₄₂N₂O₂): calcd. 438.65; obsd.439.4 [M+H]⁺.

Compound 16 was prepared in an analogous manner fromtri-methylhexamethylenediamine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min) 4.55 min. ESMS (C₃₁H₅ON₂O₂): calcd. 482.75; obsd.483.4 [M+H]⁺.

Compound 17 was prepared in an analogous manner from2,7-diaminofluorene. Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=5.2 min. ESMS (C₃₅H₄₀N₂O₂): calcd. 520.71; obsd. 521.4 [M+H]⁺.

Compound 18 was prepared in an analogous manner from4,4′-(1,3-phenylenediisopropylidene)bisaniline. Retention time (anal.HPLC: 10-70% MeCN/H₂O over 5 min)=4.73 min. ESMS (C₄₆H₅₆N₂O₂): calcd.668.96; obsd. 669.4 [M+H]⁺.

Compound 19 was prepared in an analogous manner from4,4′-diaminodiphenylmethane. Retention time (anal. HPLC: 10-90% MeCN/H₂Oover 5 min)=4.95 min. ESMS (C₃₅H₄₂N₂O₂): calcd. 522.73; obsd. 523.4[M+H]⁺.

Compound 20 was prepared in an analogous manner fromN,N-bis(3-aminoethyl)methylamine. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=3.96 min. ESMS (C₃₅H₄₂N₂O₂): calcd.441.66; obsd.442.6 [M+H]⁺.

EXAMPLE 26 Synthesis of Intermediate Compounds 2 and 200 via theFollowing Scheme

reagents and conditions: i) ethylchloroformate, EtOAc, 5N NaOH_(,) 15°C.; ii) LiAlH₄, THF, 50° C.

The compounds were synthesized from mexiletine hydrochloride inaccordance with the protocol described in Berger et. al., U.S. Pat. No.5,688,830.

Compound 2: ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.00-6.95 (d, 2H),6.90-6.84 (dd, 1H), 4.20-3.90 (m, 3H), 3.72-3.66 (d, 2H), 2.24 (s, 6H),1.34-1.30 (d, 3H), 1.28-1.20 (t, 3H).

Compound 200: ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.04-6.95 (d, 2H),6.90-6.83 (dd, 1H), 3.75-3.65 (d, 2H), 3.10-2.95 (m, 1H), 2.48 (s, 3H),2.28 (s, 6H), 1.18-1.23 (d, 3H).

EXAMPLE 27 Synthesis of Dimers of Compound 200 via the Following Scheme

reagents and conditions: i) di-halide (X—L—X), diisopropylethylamine,DMF, 90° C.

General procedure for the synthesis of compound 21 of Table A: Asolution of compound 200 (38.7 mg, 0.2 mmol) with diisopropylethylamine(54 μl, 0.3 mmol) in 200 μL of anhydrous DMF, was added to a solution of1,9-diiodononane (38.0 mg, 0.1 mmol) in 200 μL anhydrous DMF. Themixture was shaken for 20 h at 90° C., then stripped of solvent undervacuum. The resulting tarry mixture was dissolved in 1 mL of a 1:1mixture of acetonitrile and water (with 0.1% trifluoroacetic acid). Thecrude product was purified by preparative reversed phase HPLC. Retentiontime (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.58 min.; ESMS(C₃₃H₅₄N₇O₂); calcd. 510.8; obsd. 511.6 [M+H]⁺.

Compound 22 was prepared in an analogous manner fromα,α′-dibromo-p-xylene. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.94 min. ESMS (C₃₂H₄₄N₂O₂); calcd. 488.71; obsd. 489.6[M+H]⁺.

Compound 23 was prepared in an analogous manner frombis-iodoethoxyethane. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.82 min. ESMS (C₃₀H₄₈N₂O₄); calcd. 500.7; obsd. 501.4[M+H]⁺.

Compound 24 was prepared in an analogous manner from1,4-dibromo-2-butene. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.96 min. ESMS (C₂₈H₄₂N₂O₂); calcd. 438.7; obsd. 439.5[M+H]⁺.

Compound 25 was prepared in an analogous manner from(−)-trans-4,5-bis(iodomethyl)-2,2-dimethyl-1,3-dioxolane. Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.40 min. ESMS (C₃₁H₄₈N₂O₄);calcd. 512.7; obsd. 513.4 [M+H]⁺.

Compound 26 was prepared in an analogous manner from 1,10-diiododecane.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.72 min.ESMS (C₃₄H₅₆N₂O₂); calcd. 524.8; obsd. 525.4 [M+]⁺.

Compound 27 was prepared in an analogous manner from1,11-dibromoundecane. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.79 min. ESMS (C₃₅H₅₈N₂O₂); calcd. 538.9; obsd. 539.5[M+H]⁺.

Compound 28 was prepared in an analogous manner from1,12-dibromododecane. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.91 min. ESMS (C₃₆H₆₀N₂O₂); calcd. 552.9; obsd. 553.6[M+H]⁺.

Compound 29 was prepared in an analogous manner from1,16-dibromohexadecane. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.72 min. ESMS (C₄₀H₆₈N₂O₂); calcd. 609.0; obsd. 609.8 [M]⁺.

Compound 30 was prepared in an analogous manner from1,3-dibromo-2-propanol. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.18 min. ESMS (C₂₇H₄₂N₂O₃); calcd. 442.6; obsd. 443.5[M+H]⁺.

Compound 31 was prepared in an analogous manner from1,4-dichloro-2-butyne, using a catalytic amount of NaI. Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.11 min. ESMS (C₂₈H₄₀N₂O₂);calcd. 436.6; obsd. 437.5 [M+H]⁺.

Compound 32 was prepared in an analogous manner from 1,7-dibromoheptane.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.36 min.ESMS (C₃₁H₅₂N₂O₂); calcd. 482.8; obsd. 483.4 [M+H]⁺.

Compound 33 was prepared in an analogous manner from 1,8-dibromooctane.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.53 min.ESMS (C₃₂H₅₂N₂O₂); calcd. 496.8; obsd. 497.5 [M+H]⁺.

Compound 34 was prepared in an analogous manner from2,3-bis(bromomethyl)quinoxaline. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=4.84 min. ESMS (C₃₄H₄₄N₄O₂); calcd. 540.7; obsd.541.3 [M+H]⁺.

Compound 35 was prepared in an analogous manner from2,3-dibromo-1-propanol. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.22 min. ESMS (C₂₇H₄₂N₂O₃); calcd. 442.6; obsd. 443.5:[M+H]⁺.

Compound 36 was prepared in an analogous manner from3chloro-2-chloromethyl-1-propene, using a catalytic amount of NaI.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.30 min.ESMS (C₂₈H₄₂N₂O₂); calcd. 438.7; obsd. 439.3 [M+H]⁺.

Compound 37 was prepared in an analogous manner from4,5-dichloro-1,3-dioxolan-2one. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=3.16 min. ESMS (C₂₇H₃₈N₂O₅); calcd. 470.6; obsd.471.4 [M+H]⁺.

Compound 38 was prepared in an analogous manner fromalpha,alpha′-dibromo-p-xylene. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=4.30 min. ESMS (C₃₂H₄₄N₂O₂); calcd. 488.7; obsd.489.4 [M+H]⁺.

Compound 39 was prepared in an analogous manner frombis(2-chloroethoxymethane), using a catalytic amount of NaI. Retentiontime (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.13 min. ESMS(C₂₉H₄₆N₂O₄); calcd. 486.7; obsd. 487.3 [M+H]⁺.

Compound 40 was prepared in an analogous manner frombis(4-chlorobutyl)ether, using a catalytic amount of NaI. Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.26 min. ESMS (C₃₂H₅₂N₂O₃);calcd. 512.8; obsd. 513.4 [M+H]⁺.

Compound 41 was prepared in an analogous manner frombis[2-(2-chloroethoxy)ethyl]ether, using a catalytic amount of NaI.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.16 min.ESMS (C₃₂H₅₂N₂O₅); calcd. 544.8; obsd. 545.5 [M+H]⁺.

Compound 42 was prepared in an analogous manner from carbonic acidbis(2-chloroethyl)ester, using a catalytic amount of NaI. Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.13 min. ESMS (C₂₉H₄₄N₂O₅);calcd. 500.7; obsd. 501.4 [M+H]⁺.

Compound 43 was prepared in an analogous manner from ethyl2,3-dibromopropionate. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=4.78 min. ESMS (C₂₉H₄₄N₂O₄); calcd. 484.7; obsd. 485.5[M+H]⁺.

Compound 44 was prepared in an analogous manner from ethylene glycoldichloroacetate. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=4.09 min. ESMS (C₃₀H₄₄N₂O₆); calcd. 528.7; obsd. 529.3 [M+H]⁺.

Compound 45 was prepared in an analogous manner fromN,N′-bis(2-chloroethyl)oxamide, using a catalytic amount of NaI.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.96 min.ESMS (C₃₀H₄₆N₄O₄); calcd. 526.7; obsd. 527.5 [M+H]⁺.

Compound 46 was prepared in an analogous manner from 1,3-iodopropane.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.09 min.ESMS (C₂₇H₄₂N₂O₂); calcd. 426.6; obsd. 427.3 [M+H]⁺.

EXAMPLE 28 Synthesis of Compounds 3 and 300 via the Following Scheme

reagents and conditions: i) AcCl, EtOAc, 5N NaOH, 15° C.; ii) BH₃Me₂S,THF, reflux.

Compound 3A, and compound 300 were synthesized from mexiletinehydrochloride in accordance with the protocol described in Berger et.al. U.S. Pat. No. 5,688,830.

Compound 3: ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.02-6.95 (d, 2H),6.90-6.86 (dd, 1H), 4.31-4.20 (m, 1H), 3.72-3.66 (d, 2H), 2.24 (s, 6H),1.97 (s, 3H), 1.38-1.29 (d, 3H).

Compound 300: Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=3.97min. ESMS (C₁₃H₂₁NO): calcd. 207.31, obsd. 208.1 [M+H]⁺.

EXAMPLE 29 Synthesis of Dimers of Compound 300 via the Following Scheme

reagents and conditions: i) di-halide (X—L—X), diisopropylethylamine,DMF, 90° C.

General procedure for the synthesis of compound 47 of Table A: Asolution of compound 300 (41.4 mg, 0.2 mmol) with diisopropylethylamine(54 μl, 0.3 mmol) in 200 μL anhydrous DMF, was added to a solution ofbis-iodoethoxyethane (37.0 mg, 0.1 mmol) in 200 μL anhydrous DMF. Themixture was shaken for 20 h at 90° C., then stripped of solvent undervacuum. The resulting tarry mixture was dissolved in 1 mL of a 1:1mixture of acetonitrile and water (with 0.1% trifluoroacetic acid). Thecrude product was purified by preparative reversed phase HPLC. Retentiontime (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.79 min. ESMS(C₃₂H₅₂N₂O₄): calcd. 528.78, obsd. 529.4 [M++H]⁺.

Compound 48 was prepared in an analogous manner from(−)-trans-4,5-bis(iodomethyl)-2,2-dimethyl-1,3-dioxolane. Retention time(anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.33 min ESMS (C₃₃H₅₂N₂O₄):calcd. 540.79 obsd. 541.5 [M+H]⁺.

Compound 49 was prepared in an analogous manner from 1,10-diiododecane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.49 min. ESMS(C₃₆H₆₀N₂O₂): calcd. 552.88, obsd. 553.5 [M+H]⁺.

Compound 50 was prepared in an analogous manner from1,11-dibromoundecane. Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=4.61 min. ESMS (C₃₇H₆₂N₂O₂): calcd. 566.91, obsd. 567.5 [M+H]⁺.

Compound 51 was prepared in an analogous manner from1,12-dibromododecane. Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=4.72 min. ESMS (C₃₈H₆₄N₂O₂): calcd. 580.94, obsd. 581.7 [M+H]⁺.

Compound 52 was prepared in an analogous manner from1,16-dibromohexadecane. Retention time (anal. HPLC: 10-90% MeCN/H₂O over5 min)=4.90 min. ESMS (C₄₂H₇₂N₂O₂): calcd. 637.05, obsd. 637.7 [M+H]⁺.

Compound 53 was prepared in an analogous manner from 1,7-dibromoheptane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.09min. ESMS(C₃₃H₅₄N₂O₂): calcd. 510.8, obsd. 511.7 [M+H]⁺.

Compound 54 was prepared in an analogous manner fromα,α′-dibromo-m-xylene. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.05 min. ESMS (C₃₄H₄₈N₂O₂): calcd. 516.77, obsd. 517.6 [M+H]⁺.

Compound 55 was prepared in an analogous manner fromα,α′-dibromo-p-xylene. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=3.98 min. ESMS (C₃₄H₄₈N₂O₂): calcd. 516.77, obsd. 517.6 [M+H]⁺.

Compound 56 was prepared in an analogous manner frombis(2-chloroethoxymethane). Retention time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=3.80 min. ESMS (C₃₁H₅₀N₂O₄): calcd. 514.75, obsd. 515.4[M+H]⁺.

Compound 57 was prepared in an analogous manner frombis(4-chlorobutyl)ether. Retention time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=3.98 min. ESMS (C₃₄H₅₆N₂O₃): calcd. 540.83, obsd. 541.5[M+H]⁺.

Compound 58 was prepared in an analogous manner frombis[2-(2-chloroethoxy)ethyl]ether. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=3.85 min. ESMS (C₃₄H₅₆N₂O₅): calcd. 572.83, obsd.573.7 [M+H]⁺.

Compound 59 was prepared in an analogous manner from ethylene glycoldichloroacetate. Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5min)=3.74 min. ESMS (C₃₂H₄₈N₂O₆): calcd. 556.74, obsd. 557.3 [M+H]⁺.

Compound 60 was prepared in an analogous manner from 1,3-diodopropane.Retention time (anal. HPLC: 10-70% MeCN/H20 over 5 min)=3.80 min. ESMS(C₂₉H₄₆N₂O₂): calcd. 454.70, obsd. 455.3 [M+H]⁺.

EXAMPLE 30 Synthesis of Dimers of Mexiletine: Reductive Alkylation viathe Following Scheme

reagents and conditions: I) dialdehyde, EtOH, r.t., ii) NaBH₄, r.t.

General procedure for the synthesis of compound 61 of Table A: To asolution of mexiletine (neutral; 35.9 mg, 0.2 mmol) in 200 μL anhydrousethanol, was added a solution of 2,5-thiophenedicarboxaldehyde (14.0 mg,0.1 mmol) in 200 μL anhydrous ethanol. After shaking for 12 h at 25° C.,the mixture was then treated with a solution of NaBH₄ (15.2 mg, 0.4mmol) in ethanol. The final mixture was shaken for 2 h at 25° C., andthen quenched with a solution of 5% trifluoroacetic acid inacetonitrile/water (1:1). After concentration of the mixture underreduced pressure, the residue was dissolved in 1 mL of a 1:1 mixture ofacetonitrile and water (with 0.1% trifluoroacetic acid). This crudeproduct was purified by preparative reversed phase HPLC. Retention Time(anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.28 min. ESMS (C₂₈H₃₈N₂O₂S):calcd. 466.69, obsd. 467.4 [M+H]⁺.

Compound 62 was prepared in an analogous manner from2,2′-(ethylenedioxy)dibenzaldehyde. Retention Time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.60 min. ESMS (C₃₈H₄₈N₂O₄): calcd. 596.81, obsd.597.4 [M+H]⁺.

Compound 63 was prepared in an analogous manner from2,2′-(hexamethylenedioxy)dibenzaldehyde. Retention Time (anal. HPLC:10-70% MeCN/H₂O over 5 min)=4.92 min. ESMS (C₄₂H₅₆N₂O₄): calcd. 652.92,obsd. 653.4 [M+H]⁺.

Compound 64 was prepared in an analogous manner from2,2′-(trimethylenedioxy)dibenzaldehyde. Retention Time (anal. HPLC:10-70% MeCN/H₂O over 5 min)=4.75 min. ESMS (C₃₉H₅₀N₂O₄): calcd. 610.84,obsd. 611.4 [M+H]⁺.

Compound 65 was prepared in an analogous manner from2,3-thiophenedicarboxaldehyde. Retention Time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.64 min. ESMS (C₂₈H₃₈N₂O₂S): calcd. 466.69, obsd.467.4 [M+H]⁺.

Compound 66 was prepared in an analogous manner from2,6-pyridinedicarboxaldehyde. Retention Time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.31 min. ESMS (C₂₉H₃₉N₃O₂): calcd. 461.65, obsd.462.4 [M+H]⁺.

Compound 67 was prepared in an analogous manner from2-hydroxy-5-methylisophthalaldehyde. Retention Time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.59 min. ESMS (C₃₁H₄₂N₂O₃): calcd. 490.69, obsd.491.4 [M+H]⁺.

Compound 68 was prepared in an analogous manner from3,3′-(ethylenedioxy)di-p-anisaldehyde. Retention Time (anal. HPLC:10-70% MeCN/H₂O over 5 min)=4.83 min. ESMS (C₄₀H₅₂N₂O₆): calcd. 656.86,obsd. 657.6 [M+H]⁺.

Compound 69 was prepared in an analogous manner from5-formylsalicylaldehyde. Retention Time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=4.28 min. ESMS (C₃₀H₄₀N₂O₃): calcd. 476.65, obsd. 476.4[M+H]⁺.

Compound 70 was prepared in an analogous manner from6,6′-dihydroxy-5,5′-dimethoxy-(1,1′-biphenyl)-3,3′-dicarboxaldehyde.Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.50 min. ESMS(C₃₈H₄₈N₂O₆): calcd. 628.81, obsd. 629.4 [M+H]⁺.

Compound 71 was prepared i an analogous manner frombis(2-formylphenyl)ether. Retention Time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=4.66 min. ESMS (C₃₆H₄₄N₂O₃): calcd. 552.76, obsd. 553.4[M+H]⁺.

Compound 72 was prepared in an analogous manner from isophithalaldehyde.Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.39 min. ESMS(C₃₀H₄₀N₂O₂): calcd. 460.66, obsd. 461.2 [M+H]⁺.

EXAMPLE 31 Synthesis of Compound 400 via the Following Scheme

reagents and conditions: i) PPh₃, DEAD, THF, ii) TFA, CH₂Cl₂

To a stirred, cold solution of 2,6-dimethylphenol (11.25 g, 92.1 mmol),N-boc4-piperidinethanol (compound 4; 21.12 g, 92.1 mmol), andtriphenylphosphine (27.30 g, 104.1 mmol) in 400 mL THF in ice bath, wasadded dropwisely over 30 min 14.6 mL (104.1 mmol) of diethylazodicarboxylate (DEAD). The mixture was allowed to warm slowly toambient temperature and stirred for 12 h. After concentration in vacuo,the residue was treated with hexane/dichloromethane to precipitate outtriphenylphosphine oxide, which was filtered off. The filtrate wasconcentrated in vacuo, and the residue was chromatographed on silicagel. Evaporation of the appropriate fractions afforded boc-protectedether product.

Deprotection of N-Boc group of the ether product was done by using TFA.The above product (10 g) was dissolved in 300 mL of dichloromethane,cooled to 0° C., and treated with 50 mL of TFA, dropwisely over theperiod of 30 min. The mixture was warmed gradually to rt over 2 h,concentrated, and triturated with dichloromethane to give the desiredamine product as an off-white solid. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm)7.00-6.97 (d, 2H), 6.90-6.85 (dd, 1H), 3.86-3.82 (t, 2H), 3.42-3. 38 (d,2H), 3.05-2.98 (t, 3H), 2.25 (s, 6H), 2.12-1.90 (m, 3H), 1.84-1.78 (q,2H), 1.55-1.46 (m, 2H). ESMS (C₁₅H₂₃NO): calcd. 233.35; obsd. 234.2[M+H]⁺.

EXAMPLE 32 Synthesis of Dimers of Compound 400 via the Following Scheme

reagents and conditions: i) di-halide (X—L—X), diisopropylethylamine,DMF, 90° C.

Table 13 lists linkers 73-84 for dimers of structure B.

TABLE B No. Linker 73 —CH₂-Z-CH₂-where Z =(−)-trans-2,2-dimethyl-[1,3]dioxolan- 4,5-yl 74—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 75 —CH₂—CH(OH)—CH₂— 76 —(CH₂)₃— 77 —(CH₂)₆—78 —CH₂—C(CH₂)—CH₂— 79 —CH₂-Z-CH₂— where Z = 1,3-phenyl 80 —CH₂-Z-CH₂—where Z = 1,4-phenyl 81 —(CH₂)₄—O—(CH₂)₄— 82—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 83 —(CH₂)₂—NH—C(O)—C(O)—NH—(CH₂)₂—84 —CH₂-Z-CH₂— where Z = 4,4′-biphenyl

General procedure for synthesis of compound 75 of Table B: A solution ofcompound 400 (TFA salt; 69.5 mg, 0.2 mmol) with diisopropylethylamine(108 μl, 0.6 mmol) in 250 μL of anhydrous DMF, was added to a solutionof 1,3-diiodo-2-propanol (31.8 mg, 0.1 mmol) in 250 μL of anhydrous DMF.The mixture was shaken for 20 h at 90° C., and concentrated undervacuum, yielding tarry mixture. It was dissolved in 1 mL of 50% aqueousacetonitrile (with 0.1% trifluoroacetic acid), and purified by reversedphase preparative HPLC. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.03 min. ESMS (C₃₃H₅₀N₂O₃): calcd. 522.78; obsd. 523.6 [M+H]⁺.

Compound 73 was prepared in an analogous manner from(−)-trans-4,5-bis(iodomethyl)-2,2-dimethyl-1,3-dioxolane. Retention time(anal. HPLC: 10-90% MeCN/H₂O over 5 min)=4.40 min. ESMS (C₃₇H₅₆N₂O₄):calcd. 592.86; obsd. 593.4 [M+H]⁺.

Compound 74 was prepared in an analogous manner frombis-iodoethoxyeffiane. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.13 min. ESMS (C₃₆H₅₆N₂O₄): calcd. 580.85; obsd. 581.6 [M+H]⁺.

Compound 76 was prepared in an analogous manner from 1,3-diiodopropane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.05 min. ESMS(C₃₃H₅₀N₂O₂): calcd. 506.77; obsd. 507.4 [M+H]⁺.

Compound 77 was prepared in an analogous manner from 1,6-diiodohexane.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.26 min. ESMS(C₃₆H₅₆N₂O₂): calcd. 548.85; obsd. 549.6 [M+H]⁺.

Compound 78 was prepared in an analogous manner from3-chloro-2-chloromethyl-1-propene. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.22 min. ESMS (C₃₄H₅₀N₂O₂): calcd. 518.78; obsd.519.6 [M+H]⁺.

Compound 79 was prepared in an analogous manner fromα,α′-dibromo-m-xylene. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.34 min. ESMS (C₃₈H₅₂N₂O₂): calcd. 568.84; obsd. 569.4 [M+H]⁺.

Compound 80 was prepared in an analogous manner fromα,α′-dibromo-p-xylene. Retention time (anal. HPLC: 10-70% MeCN/H₂O over5 min)=4.28 min. ESMS (C₃₈H₅₂N₂O₂) calcd. 568.84; obsd. 569.4 [M+H]⁺.

Compound 81 was prepared in an analogous manner frombis(4-chlorobutyl)ether. Retention time (anal. HPLC: 10-70% MeCN/H₂Oover 5 min)=4.28 min. ESMS (C₃₈H₆₀N₂O₃): calcd. 592.91; obsd. 593.4[M+H]⁺.

Compound 82 was prepared in an analogous manner frombis[2-(2-chloroethoxy)ethyl]ether. Retention Time (anal. HPLC: 10-70%.MeCN/H₂O over 5 min)=4.14 min. ESMS (C₃₈H₆₀N₂O₃): calcd. 624.90; obsd.625.6 [M+H]⁺.

Compound 83 was prepared in an analogous manner fromN,N′-bis(2-chloroethyl)oxamide. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.08 min. ESMS (C₃₆H₅₄N₄O₄): calcd. 606.84; obsd.607.6 [M+H]⁺.

Compound 84 was prepared in an analogous manner from4,4-bis(chloromethyl)1,1-biphenyl. Retention time (anal. HPLC: 10-70%MeCN/H₂O over 5 min)=4.63 min. ESMS (C₄₄H₅₆N₂O₂): calcd. 644.94; obsd.645.6 [M+H]⁺.

EXAMPLE 33 Synthesis of Compound 1 of Table C via the Following Scheme

reagents and conditions: i) 1,2-bis-(2-iodoethoxy)ethane, Na₂CO₃, NaI,CH₃CN, 150°° C.

A suspension of acetonitrile (14 mL) containing Table C, compound 1 ofTable A (43.4 mg, 0.092 mmol), 1,2-bis(2-iodoethoxy)ethane (21 μL, 0.115mmol), potassium carbonate (63.6 mg, 0.46 mmol), and potassium iodide(7.6 mg, 0.046 mmol) in a thick-walled, sealed tube was heated at 150°C. for 12 h. The reaction mixture was cooled, filtered, and concentratedin vacuo. The resulting residue was dissolved in 1 mL of 50% aqueousacetonitrile (with 0.1% trifluoroacetic acid), and purified by reversedphase preparative HPLC. Retention time (anal. HPLC: 2-90% MeCN/H₂O over5 min)=2.96 min (a mixture of diastereomers). ESMS (C₃₄H₅₄N₂O₆): calcd.586.8; obsd. 587.4 [M+H]⁺.

TABLE C No. Molecule 1

2

3

4

5

Table C lists linkers that can be employed in the above scheme.

EXAMPLE 34 Synthesis of Dimers via the Following Scheme

reagents and conditions: i) di-halide (X—L—X), diisopropylethylamine,DMF, 90° C.

Compound 100 was prepared in racemic form following procedures asdescribed in L. A. Flippin, et al., EP0869119 A1.

Table D lists linkers 86-103 for dimer D.

TABLE D No. Linker 86 —CH₂-Z-CH₂— where Z = 1,4-phenyl 87 —(CH₂)₉— 88—CH₂—CH═CH—CH₂— (trans isomer) 89 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 90—CH₂-Z-CH₂— where Z = (−)-trans-2,2-dimethyl-[1,3] dioxolan-4,5-yl 91—(CH₂)₁₀— 92 —(CH₂)₁₁— 93 —(CH₂)₁₂— 94 —(CH₂)₁₆— 95 —CH₂—CH(OH)—CH₂— 96—CH₂—C═C—CH₂— 97 —(CH₂)₇— 98 —(CH₂)₈— 99 —CH₂-Z-CH₂— where Z =2,3-quinoxalinyl 100 —CH₂—CH(CO₂H)— 101 —CH₂—CH(CH₃)—CH₂— 102 —CH₂-Z-CH₂where Z = 1,3-phenyl 103 —(CH₂)₂—O—CH₂—O—(CH₂)₂—

General procedure for the synthesis of compound 86 of Table D: Asolution of compound 100 (43.8 mg, 0.2 mmol) with diisopropylethylamine(54 μl, 0.3 mmol) in 200 μL of anhydrous DMF, was added to a solution ofα,α′-dibromo-p-xylene (0.1 mmol) in 200 μL anhydrous DMF. The mixturewas shaken for 20 h at 90° C., then stripped of solvent under vacuum.The resulting tarry mixture was dissolved in 1 mL of a 1:1 mixture ofacetonitrile and water (with 0.1% trifluoroacetic acid). The crudeproduct was purified by preparative reversed phase HPLC to afford 26.2mg of the desired material as the TFA salt. ESMS (C₃₆H₄₈N₂O₂): calcd.540.79; obsd. 541 [M+H]⁺. Retention time (anal. HPLC: 2-90% MeCN/H₂Oover 5 min)=4.7 min.

Compound 87 was prepared with 1,9-dibromononane (0.1 mmol) using thestandard procedure to provide 16.3 mg of the desired material as the TFAsalt. ESMS (C₃₇H₅₈N₂O₂): calcd. 562.8; obsd. 563 [M+H]⁺. Retention time(anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.8 min.

Compound 88 was prepared with 1,4-dibromo-2-butene (0.1 mmol) using thestandard procedure to provide the desired material as the TFA salt. ESMS(C₃₂H₄₆N₂O₂): calcd. 490.72; obsd. 491 [M+H]⁺. Retention time (anal.HPLC: 2-90% MeCN/H₂O over 5 min)=4.76 min.

Compound 89 was prepared with 1,2-bis(2-iodoethoxy)ethane (0.1 mmol)using the standard procedure to provide 18.7 mg of the desired materialas the TFA salt. ESMS (C₃₄H₅₂N₂O₄): calcd. 552.80; obsd. 553 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.7 min.

Compound 90 was prepared with(−)-trans4,5-bis(idodomethyl))-2,2-dimethyl-1,3-dioxolane (0.1 mmol)using the standard procedure to provide 8.5 mg of the desired materialas the TFA salt. ESMS (C₃₅H₅₂N₂O₄): calcd. 564.81; 565.3 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.8 min.

Compound 91 was prepared with 1,10-diiododecane (0.1 mmol) using thestandard procedure to provide 10.5 mg of the desired material as the TFAsalt. ESMS (C₃₈H₆₀N₂O₂): calcd. 576.91; obsd. 577.6 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.4 min.

Compound 92 was prepared with 1,11-dibromoundecane (0.1 mmol) using thestandard procedure to provide 11.4 mg of the desired material as the TFAsalt. ESMS (C₃₉H₆₂N₂O₂): calcd. 590.93; obsd. 591.5 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.49 min.

Compound 93 was prepared with 1,12-dibromododecane (0.1 mmol) using thestandard procedure to provide 13.2 mg of the desired material as the TFAsalt. ESMS (C₄₀H₆₄N₂O₂): calcd. 604.96; obsd. 605.6 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.58 min.

Compound 94 was prepared with 1,16-dibromohexadecane (0.1 mmol) usingthe standard procedure to provide 10.9 mg of the desired material as theTFA salt. ESMS (C₄₄H₇₂N₂O₂): calcd. 661.07; obsd. 661.7 [M]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.93 min.

Compound 95 was prepared with 1,3-dibromo-2-propanol (0.1 mmol) usingthe standard procedure to provide 7.5 mg of the desired material as theTFA salt. ESMS (C₃₁H₄₆N₂O₃): calcd. 494.72; obsd. 495.4 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=3.95 min.

Compound 96 was prepared with 1,4-dichloro-2-butyne (0.1 mmol) using thestandard procedure to provide 11.4 mg of the desired material as the TFAsalt. ESMS (C₃₂H₄₄N₂O₂): calcd. 488.71; obsd. 489.4 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=3.97 min.

Compound 97 was prepared with 1,7-dibromoheptane (0.1 mmol) using thestandard procedure to provide 12.7 mg of the desired material as the TFAsalt. ESMS (C₃₅H₅₄N₂O₂): calcd. 534.83; obsd. 535.5 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.16min.

Compound 98 was prepared with 1,8-diiododecane (0.1 mmol) using thestandard procedure to provide 16.3 mg of the desired material as the TFAsalt. ESMS (C₃₆H₅₆N₂O₂) 548.85; obsd. 549.4 [M+H]⁺. Retention time(anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.22 min.

Compound 99 was prepared with 2,3-bis(bromomethyl)quinoxaline (0.1 mmol)using the standard procedure to provide the desired material as the TFAsalt. ESMS (C₃₈H₄₈N₄O₂): calcd. 592.82; obsd. 593.6 [M+H]⁺. Retentiontime (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.19 min.

Compound 100 was prepared with 2,3-dibromopropionic acid (0.1 mmol)using the standard procedure to provide 10.4 mg of the desired materialas the TFA salt. ESMS (C₃₁H₄₄N₂O₄): calcd. 508.70; obsd. 509.5 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over win)=4.23 min.

Compound 101 was prepared with 3chloro-2-chloromethyl-1-propene (0.1mmol) using the standard procedure to provide 22.1 mg of the desiredmaterial as the TFA salt. ESMS (C₃₂H₄₆N₂O₂): calcd. 490.73; obsd. 491.4[M+H]⁺. Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.12 min.

Compound 102 was prepared with α,α′-dibromo-m-xylene (0.1 mmol) usingthe standard procedure to provide 22.5 mg of the desired material as theTFA salt. ESMS (C₃₆H₄₈N₂O₂): calcd. 540.79; obsd. 541.4 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=4.12 min.

Compound 103 was prepared with bis(2-chloroethoxyethane) (0.1 mmol)using the standard procedure to provide 15.4 mg of the desired materialas the TFA salt. ESMS (C₃₃H₅₀N₂O₄): calcd. 538.77; obsd. 539.4 [M+H]⁺.Retention time (anal. HPLC: 2-90% MeCN/H₂O over 5 min)=3.97 min.

EXAMPLE 35 Synthesis of Intermediate Compounds 9 to 12 via the FollowingScheme

reagents and conditions i) NaBH₄, MeOH, 0° C.; ii) methanesulfonylchloride, Et₃N, DMAP, toluene, 0° C.; iii) KCN, Bu₄NHSO₄, toluene water,rt; iv) FCH₂CO₂Et, NaOMe, MeOH, rt; v) Et-I, K₂CO₃, DMF, 75° C.

Compounds 9 to 12 were synthesized according to procedures described inNobbs, et al., WO 97/09317 and Miller, et al., EP 0 372 934 B1.

Compound 9: To solution of MeOH (150 mL) containing2,3-dichlorobenzaldehyde (20 g, 0.11 mole), cooled in ice bath, wasadded a solution of NaBH₄ (4.54 g, 0.12 mole) in 0.2 M NaOH (15 mL)slowly over 15 min under nitrogen atmosphere. After completion of theaddition, the mixture was allowed to warm up gradually to rt, andstirred for 2.5 h at rt. The reaction was quenched by cooling themixture and pouring it slowly to ice water (˜700 mL). The mixing led toformation of white precipitates. After stirring for 30 min, theprecipitates were collected on Bòchner funnel; and the collected solidwas dissolved in EtOAc (200 mL). The organic solution was washed with0.1 M NaOH (150 mL), dried over Na₂SO₄, and concentrated to dryness invacuo, yielding 2,3-dichlorobenzylalcohol as white solid (19.57 g, 97%).R_(f)=0.46 in EtOAc/hexane (1/3). ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm)7.52-7.49 (dd, 1H), 7.45-7.42 (dd, 1H), 7.33-7.27 (t, 1H), 4.70 (s, 2H).Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.00 min.

Compound 10: To a solution of toluene (100 mL) containing2,3-dichlorobenzyl alcohol (18 g, 0.102 mole), Et₃N (12.4 g, 0.123mole), and 4-dimethylaminopyridine (0.621 g, 5.08 mmole), cooled in icebath, was added methanesulfonyl chloride (14 g, 0.122 mole) whilestirring the mixture vigorously. The mixture was shaken at 0° C. for 1h, prior to allowing it left in refrigerator over 2 days. The mixturewas then diluted with EtOAc (100 mL) and brine (200 mL). After shakingin a separatory funnel, the organic phase was separated, and washed withsat. NaHCO₃ solution. Organic solution was dried over Na₂SO₄, andconcentrated in vacuo to ˜50 mL which was used directly in next stepwithout further purification. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm)7.58-7.52 (dd, 1H), 7.51-7.48 (dd, 1H), 7.36-7.30(t, 1H), 5.36 (s, 2H),3.14 (s, 3H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=4.43 min.

Compound 11: Compound 10 (2,3-dichlorobenzyl mesylate), prepared abovein toluene (˜50 mL), was diluted to ˜70 mL with toluene, and followed byaddition of Bu₄NHSO₄ (7.0 g, 0.02 mole) in water (30 mL) and KCN (10 g,0.15 mole) in water (40 mL). The mixture was stirred for 24 h at rt, anddiluted with EtOAc (200 mL) prior to washing with 0.1 M NaOH (200 mL).The organic phase was washed again with 0.1 M NaOH and brine, dried overNa₂SO₄, and concentrated in vacuo, yielding a dark brown oily residue.The crude product was purified by flash silica column chromatography byeluting with hexane/EtOAc (3/1). The product was obtained as white topale beige solid (16.35 g; 86% over two steps). R_(f)=0.58 inEtOAc/hexane (1/3). ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.56-7.53 (dd,1H), 7.50-7.47 (dd, 1H), 7.36-7.31 (t, 1H), 4.03 (s, 2H). Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.53 min.

Compound 12: To stirred suspension of compound 11 (16.6 g, 0.089 mole)in MeOH (100 mL) in water bath (rt) was added slowly NaOMe (12.05 g,0.22 mole). After stirring for 10 min, ethyl fluoroacetate (11.36 g,0.107 mole) was added to the mixture. The final mixture was stirred for24 h at it. Reaction mixture was concentrated to oily residue, and thematerial was dissolved in water (150 mL). The aqueous layer was washedwith ethyl ether (3×200 mL); white insoluble material was saved andpooled with the aqueous layer. The combined solution was acidified to pH3 by using 6M HCl, and the aqueous solution was in extracted with ethylether (500 mL). The organic solution was dried over Na₂SO₄, andconcentrated to yield product(4-fluoro-3-oxo-2-(2,3-dichlorophenyl)-butyronitrile) as pale yellowoily residue which slowly solidified. The crude product (11.29 g; 51.3%)was used in next step. R_(f)=0.76 in 5% MeOH/EtOAc/hexane (1/1). ¹H-NMR(CD₃OD, 299.96 MHz): δ (ppm) 7.57-7.54 (dd, 1H), 7.48-7.43 (dd, 1H),7.35-7.32 (t, 1H), 5.36 & 5.20 (two s, 2H).

To a solution of DMF (200 mL) containing the above nitrite (11.2 g,0.046 mole) was added K₂CO₃ (12.6 g, 0.091 mole) and ethyl iodide (14.2g, 0.091 mole). The mixture was stirred overnight at 75° C. The reactionmixture was cooled down to rt, and diluted with EtOAc (500 mL) andfollowed by addition of water (300 mL). After shaking in a separatoryfunnel, the organic phase was separated and washed with brine solution(200 mL), dried over Na₂SO₄, and concentrated to oily residue. It waspurified by flash silica column chromatography by eluting withhexane/EtOAc (4/1 to 2/1) to afford compound 12 as pale yellow oil(12.47 g; 49.1%). R_(f)=0.39 in 5% MeOH/EtOAc/hexane (1/1). ¹H-NMR(CD₃OD, 299.96 MHz): δ (ppm) 7.59-7.56 (t, 1H), 7.35-7.33 (m, 2H), 5.54& 5.39 (two s, 2H), 4.25-4.18 (q, 2H), 1.22-1.17 (t, 3H).

EXAMPLE 36 Synthesis of bis-guanidines via the Following Scheme

General procedure for the synthesis of compound 7 of Table E: To asolution of 1,5-diaminohexane (0.78 g, 7.63 mmole) in water (10 mL) wasadded methylmercaptocarboxamidine hydrogen iodide salt (5.0 g; 0.023mole) at rt. The mixture was stirred, and heated at 90° C. for 24 h inwell-ventilated hood. The reaction mixture was concentrated in vacuo,and white crystalline solid was obtained. It was suspended in 30 mL ofi-PrOH/ether (1/1), collected on Büchner funnel, and further washed with30 mL of i-PrOH/ether (1/1). The product was obtained as white solid(1.8 g; 52%) as hydrogen iodide salt. ¹H-NMR(D₂O, 299.96 MHz): δ (ppm)3.03-3.09 (t, 4H), 1.49 (m,4H), 1.27 (m, 2H).

Table E lists linkers 1-18 for dimer E.

TABLE E No. Linker R 1

— 2 —(CH₂)₃— Me 3 —CH₂—CH═CH—CH₂— (trans isomer) Me 4 —(CH₂)₆— Me 5—(CH₂)₃— H 6 —CH₂—Z—CH₂— where Z = 1,4-phenyl H 7 —(CH₂)₅— H 8 —(CH₂)₈—H 9 —(CH₂)₇— H 10 —(CH₂)₃—O—(CH₂)₄—O—(CH₂)₃— H 11 —(CH₂)₂—(CH₂)₂— H 12—(CH₂)₁₀— H 13 —(CH₂)₄— H 14 —CH₂—Z—CH₂— where Z = 1,3-phenyl H 15

H 16 —(CH₂)₃—O—(CH₂)₃— H 17 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— H 18—(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— H

Bis-guanidines below were prepared from corresponding di-amine's in ananalogous manner as described above.

Compound 1: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 4.64 (s).

Compound 2: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.30-3.25 t, 4H), 2.92 (s,6H), 1.94-1.92 (q, 2H).

Compound 3: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 5.59 (s, 2H), 3.92 (s,4H), 2.94 (s, 6H).

Compound 4: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.22-3.18 (t, 4H), 2.89(s, 6H), 1.55-1.46 (bra quin, 4H), 1.25-1.18 (bra quin, 4H).

Compound 5: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.18-3.1 (t, 4H), 1.8-1.7(quin, 2H).

Compound 6: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 7.30 (s, 4H), 4.36 (s,4H).

Compound 8: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.04-3.08 (t, 4H), 1.47(m, 4H), 1.23 (m, 8H).

Compound 9: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.08-3.04 (t, 4H),1.46-1.42 (t, 4H), 1.23 (m, 6H).

Compound 10: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.50-3.42 (m, 8H),3.20-3.14 (t, 4H), 1.80-1.70 (quin, 4H), 1.55-1.48 m, 4H).,

Compound 11: ¹H-NMR (D₂O, 299.96 MHz) δ (ppm) 3.35-3.31 (t, 4H), 2.75(m, 4H), 2.35 (s, 3H).

Compound 12: 1H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.02-3.06 (t, 4H), 1.45(m, 4H), 1.18 (m, 12H).

Compound 13: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.13 (t, 4H), 1.56 (t,4H).

Compound 14: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 7.13-7.15 (d, 2H), 7.18(s, 1H), 7.27-7.32 (t, 1H), 4.30 (s, 4H).

Compound 15: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.09-3.13 (t, 4H), 2.56(m, 8H), 2.40-2.43 (t, 4H), 1.65-1.74 (quin, 4H).

Compound 16: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.45-3.49 (t, 4H),3.14-3.18 (t, 4H), 1.71-1.79 (quin, 4H).

Compound 17: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.61 (s, 4H), 3.58-3.62(t, 4H), 3.28-3.32 (t, 4H).

Compound 18: ¹H-NMR (D₂O, 299.96 MHz): δ (ppm) 3.56 (s, 8H), 3.47-3.51(t, 4H), 3.14-3.18 (t, 4H), 1.70-1.78 (quin, 4H).

EXAMPLE 37 Synthesis of Homodimers of 5-arylpyrimidine via the FollowingScheme

General procedure for the synthesis of compound 15 of Table F: R═Me,L═(CH₂)₃): To Table E, compound 2 (0.3 g, 0.678 mmole) in a vial wasadded NaOMe (100 mg/mL; 0.77 mL; 1.43 mmole) and compound 12 (500 mg/LMeOH; 1.116 mL, 2.04 mmole). The reaction vessel was well sealed with ascrew cap, and shaken at 80° C. for 6 h. The mixture was thenconcentrated to dryness, and the dark brown residue was dissolved in 5mL of 50% aqueous acetonitrile (with 5% TFA). After filtration, thecrude product was purified by preparative reversed phase HPLC: 20 to 60%aqueous acetonitrile (0.1% TFA) over 40 min (linear gradient); 30mL/min; 254 nm. The desired product was obtained as white solid (70 mg).ESMS (C₂₈H₃₀N₈O₂F₂Cl₄); calcd. 658.41; obsd. 659.3 [M+H]⁺. ¹H-NMR(CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.70 (dd, 2H), 7.50-7.41 (dt, 2H),4.32-7.29 (dt, 2H), 5.2-5.08 (q, 4H), 3.9-3.78 (m, 4H), 3.30 (s, 6H),2.2-2.08 (quin, 2H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.63 min.

Table F lists linkers 1-18 for tile following dimer.

TABLE F No. Linker R R^(5a) R^(6a) R^(5b) R^(6b) R^(4a) R^(4b) 1—(CH₂)₃— H Cl Cl Cl Cl —CH₂F —CH₂F 2 —CH₂—Z—CH₂— where Z = 1,4-phenyl HCl Cl Cl Cl —CH₂F —CH₂F 3 —(CH₂)₅— H Cl Cl Cl Cl —CH₂F —CH₂F 4 —(CH₂)₈—H Cl Cl Cl Cl —CH₂F —CH₂F 5 —(CH₂)₇— H Cl Cl Cl Cl —CH₂F —CH₂F 6—(CH₂)₃—O—(CH₂)₄—O—(CH₂)₃— H Cl Cl Cl Cl —CH₂F —CH₂F 7—(CH₂)₂—N(CH₃)—(CH₂)₂— H Cl Cl Cl Cl —CH₂F —CH₂F 8 —(CH₂)₁₀— H Cl Cl ClCl —CH₂F —CH₂F 9 —(CH₂)₄— H Cl Cl Cl Cl —CH₂F —CH₂F 10 —CH₂—Z—CH₂— whereZ = 1,3-phenyl H Cl Cl Cl Cl —CH₂F —CH₂F 11

H Cl Cl Cl Cl —CH₂F —CH₂F 12 —(CH₂)₃—O—(CH₂)₃— H Cl Cl Cl Cl —CH₂F —CH₂F13 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— H Cl Cl Cl Cl —CH₂F —CH₂F 14—(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— H Cl Cl Cl Cl —CH₂F —CH₂F 15—N(CH₃)—(CH₂)₃—N(CH₃)— Me Cl Cl Cl Cl —CH₂F —CH₂F 16—N(CH₃)—CH₂—CH═CH—CH₂—N(CH₃)— Me Cl Cl Cl Cl —CH₂F —CH₂F (trans isomer)17 —N(CH₃)—(CH₂)₆—N(CH₃)— Me Cl Cl Cl Cl —CH₂F —CH₂F 18

— Cl Cl Cl Cl —CH₂F —CH₂F

Compound 1 was synthesized from Table E, compound 5 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.14. ESMS (C₂₅H₂₂Cl₄F₂N₈); calcd. 642.37; obsd. 643.1 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (d, 4H), 7.46-7.42 (t,2H), 7.33-7.31 (d, 4H), 4.98 (s, 2H), 4.82 (s, 2H), 4.30-4.24 (t, 4H),2.38-2.36 (m, 2H).

Compound 2 was prepared from Table E, compound 6 in an analogous way asdescribed above. ESMS (C₃₀H₂₄N₈F₂Cl₄); calcd. 676.4; obsd. 677.1 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (dd, 2H), 7.50-7.42 (dt,2H), 7.38-7.35 (dd, 2H), 7.34 (d, 4H), 5.49 (s, 4H), 5.03 & 4.88 (two s,4H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.48min.

Compound 3 was synthesized from Table E, compound 7 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.02. ESMS (C₂₇H₂₆Cl₄F₂N₈); calcd. 642.37; obsd. 643.1 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.70 (d, 4H), 7.48-7.42 (t,2H), 7.33-7.30 (d, 4H), 4.97 (s, 2H), 4.81 (s, 2H), 4.18-4.13 (t, 4H),1.92-1.87 (m, 4H), 1.64-1.62 (m, 2H).

Compound 4 was synthesized from Table E, compound 8 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.24. ESMS (C₃₀H₃₂Cl₄F₂N₈); calcd. 684.45; obsd. 685.4 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (d, 4H), 7.47-7.42 (t,2H), 7.33-7.30 (d, 4H), 4.96 (s, 21I), 4.81 (s, 2H₁), 4.12-4.07 (t, 4H),1.81 (m, 4H), 1.46-1.44(m, 8H).

Compound 5 was prepared from Table E, compound 9 in an analogous way asdescribed above. ESMS (C₂₉H₃₀N₈F₂Cl₄); calcd. 670;4; obsd. 671.1 [M+H]⁺.¹H-NMR (CD₃OD, 299.96. MHz): δ (ppm) 7.72-7.69 (dd, 2H), 7.48-7.42 (t,2H), 7.33-7.30 (dd, 2H), 4.96 & 4.81 (two s, 4H), 4.14-4.09 (t, 4H),1.81 (br, 4H), 1.48 (br,6H). Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=3.58 min.

Compound 6 was prepared from Table E, compound 10 in an analogous way asdescribed above. ESMS (C₃₂H₃₆N₈F₂Cl₄O); calcd. 744.5; obsd. 745.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.69 (dd, 2H),7.48-7.42 (t, 2H), 7.33-7.30 (dd, 2H), 4.97 & 4.83 (two d, 4H),4.23-4.19 (t, 4H), 3.61-3.57 (t, 4H0, 3.48 (br s, 4H), 2.15-2.11 (br m,4H), 1.61 (br s,

Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.2 min.

Compound 7 was prepared from Table E, compound 11 in an analogous way asdescribed above. ESMS (C₂₇H₂₇N₉F₂Cl₄); calcd. 657.4; obsd. 658.2 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (dd, 2H), 7.48-7.43 (t,2H), 7.36-7.33 (m, 2H), 4.99 & 4.83 (two s, 4H), 4.37-4.33 (t, 41),3.10-3.0 (m, 4H), 2.56 (s, 3H). Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=3.38 min.

Compound 8 was synthesized from Table E, compound 12 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.46. ESMS (C₃₂H₃₆Cl₄F₂N₈); calcd. 712.50; obsd. 713.3 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (d, 4H), 7.47-7.42 (t,2H), 7.33-7.30 (d, 4H), 4.96 (s, 2H), 4.80 (s, 2H), 4.12-4.06 (t, 4H),1.79 (m, 4H), 1.36 (m, 4H).

Compound 9 was synthesized from Table E, compound 13 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=2.85. ESMS (C₂₆H₂₄Cl₄F₂N₈); calcd. 628.34; obsd. 629.0 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.69 (d, 4H), 7.48-7.43 (t,2H), 7.34-7.32 (d, 4H), 4.97 (s, 2H), 4.81 (s, 2H), 4.18 (m, 4H), 1.99(m, 4H).

Compound 10 was synthesized from Table E, compound 14 in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.03. ESMS (C₃₀H₂₄Cl₄F₂N₈); calcd. 676.38; obsd. 677.0[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.72 (d, 4H), 7.59-7.54(t, 1H), 7.47-7.44 (t, 2H), 7.39-7.37 (d, 2H), 7.32-7.30 (d, 4H), 6.94(s, 1H), 5.51 (s 4H), 5.03 (s, 2H), 4.88 (s, 2H).

Compound 11 was synthesized from Table E, compound 15 in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.66. ESMS (C₃₂H₃₆Cl₄F₂N₁₀); calcd. 740.51; obsd. 741.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.69 (d, 4H), 7.48-7.42(t, 2H), 7.31-7.30 (d, 4H), 4.97 (s, 2H), 4.82 (s, 2H), 4.20-4.15 (t14H), 3.11 (m, 8H), 2.21-2.16 (t, 4H), 1.87-1.84 (quin, 4H).

Compound 12 was synthesized from Table E, compound 16 in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=1.44. ESMS (C₂₈H₂₈Cl₄F₂N₈O); calcd. 672.39; obsd. 673.1[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (d, 4H), 7.48-7.42(t, 2H), 7.33-7.30 (d, 4H),:4.97 (s, 2H), 4.82 (s, 2H), 4.23-4.19 (t,4H), 3.74-3.70 (t, 4H), 3.64-3.60 (t, 2H), 2.16-2.08 (p, 2H),1.88-1.84(p, 2H).

Compound 13 was synthesized from Table E, compound 17 in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.41. ESMS. (C₂₈H₂₈Cl₄F₂N₈O₂); calcd. 688.39; obsd. 689.3[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (d, 4H), 7.48-7.43(t, 2H), 7.33-7.31 (d, 4H), 4.99 (s, 2H), 4.84 (s, 2H), 4.39-4.36 (t,4H), 3.92-3.89 (t, 4H), 3.69 (s, 4H).

Compound 14 was prepared from Table E, compound in an analogous way asdescribed above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=1.49. ESMS (C₃₂H₃₆Cl₄F₂N₈O₃); calcd. 760.50; obsd. 761.2 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (d, 4H), 7.48-7.42 (t,2H), 7.33-7.31 (d, 4H), 4.96 (s, 2H), 4.81 (s, 2H), 4.24-4.20 (t, 4H),3.74-3.70 (t, 2H), 3.65 (s, 8H), 3.69-3.61 (t, 2H), 2.20-2.12 (p, 2H),1.87-1.84 (quin, 2H).

Compound 16 was prepared from Table E, compound 27 in an analogous wayas described above. ESMS (C₂₉H₃₀N8F₂Cl₄); calcd. 670.4; obsd. 671.3[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.74-7.71 (dd, 2H),7.49-7.44 (t, 2H), 7.33-7.31 (d, 2H), 5.86 (s, 2H), 5.20-5.10 (dd, 2H),5.02-4.95 (dd, 2H), 4.48-4.35 (brt, 4H), 4.10 (s, 8H), 3.25 (s, 6H).Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.68 min.

Compound 17 was synthesized from Table E, compound 4 in an analogous wayas described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=3.51. ESMS (C₃₀H₃₂Cl₄F₂N₈); calcd. 684.44; obsd. 685.5 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.74-7.72 (d, 4H), 7.48-7.44 (t,2H), 7.33-7.31 (d, 4H), 5.11 (m, 2H), 4.96 (m, 2H), 3.74 (m, 4H), 3.26(s, 6H), 1.74 (m, 4H), 1.46 (m, 4H).

Compound 18 of Table F was prepared from Table E, compound 1 in ananalogous way as described above. ESMS (C₂₆H₂₂N₈F₂Cl₄); calcd. 626.3;obsd. 627.2 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.75-7.72 (dd,2H), 7.50-7.45 (t, 2H), 7.35-7.32 (dd, 2H), 5.21-5.10 (dd, 2H),5.05-4.98 (dd, 2H), 4.10 (s, 8H). Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=3.82 min.

EXAMPLE 38 Synthesis of Mexiletine-guaidines via the Following Scheme

reagents and conditions: i) H₂N—L—NH₂, THF, rt; ii)1-H-pyrazole-1-[N,N′-bis(tert-butoxycarbonyl)]carboxamidine, rt; iii)NaCNBH₃, rt; iv) TFA, CH₂Cl₂, 0° C.

General procedure for the synthesis of compound 1 of Table G: L=(CH₂)₆):(Step i): To a solution of 1,6-diaminohexane in TBF (10 mL) was, added2,6-dimethylphenyloxyacetone (compound 1; 1.0 g, 5.6 mmole) in THF (5mL). After stirring at rt for 1.5 h, mass spectrometric analysis of thereaction mixture indicated the formation of imine as a major species:ESMS (C₁₇H₂₈N₂O): calcd. 276.4; obsd. 277.3 [M+H]⁺.

Table G lists linkers 1-15 for dimer G.

TABLE G No. Linker 1 —(CH₂)₆— 2

3 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 4 —(CH₂)₅— 5—(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— 6 —(CH₂)₂—N(CH₃)—(CH₂)₂— 7—(CH₂)₃—O—(CH₂)₃— 8

9 —CH₂—Z—CH₂— where Z = 1,3-phenyl 10 —(CH₂)₂—S—(CH₂)₂— 11 —CH₂—Z—CH₂—where Z = 1,4-phenyl 12 —(CH₂)₄— 13 —(CH₂)₂—O—(CH₂)₂— 14 —(CH₂)₂— 15

(Step ii): To the imine solution was added1-H-pyrazole-1-[N,N′-bis(tert-butoxycarbonyl)carboxamidine] (2.10 g,6.77 mole). The mixture was stirred at rt for 3 h. Again, mass analysisof the reaction mixture indicated formation of N,N′-bis-Boc-protectedguanidine (ESMS: obsd. 519.5 [M+H]⁺).

(Step iii): The mixture was then treated with NaCNBH₃ (0.423 g, 6.73mmole) to reduce the imine functionality to amine. After stirring at rtor 1 h, the reaction was quenched by addition of water (1 mL). Thereaction mixture was diluted with EtOAc (100 mL), washed with 0.1 MNaOH/brine solution, and dried over Na₂SO₄. The organic solution wasevaporated in vacuo, yielding amine product as white solid. This crudeproduct was used in next step without further purification. ESMS: calcd.520.8; Obsd. 522.2 [M+H]⁺.

(Step iv): The above product was dissolved in CH₂Cl₂ (15 mL), and thenfollowed by addition of TFA (8 mL). The mixture was stirred overnight,and concentrated in vacuo, yielding colorless oily residue. The crudeproduct was dissolved in 50% aqueous acetonitrile, and purified bypreparative reversed phase HPLC: 10 to 40% aq. MeCN over 40 min (lineargradient); 40 mL/min; 214 nm. The product was obtained as colorless oil(500 mg; overall 16% to four steps). ESMS (C₁₈H₃₂N₄O₁); calcd. 320.5;obsd. 321.4 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.04-7.0 (d,2H), 6.98-6.96 (t, 1H), 4.0-3.95 (m, 2H), 3.78-3.68 (m, 1H), 3.2-3.17(m, 4H), 2.29 (s, 6H), 1.82-1.7 (br m, 2H), 1.64-1.58 (br m, 2H),1.57-1.4 (m, 7H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=2.01 min.

Compound 2 was prepared from N,N′-bis(3-amino-1-propyl)piperazine in ananalogous manner as described above. ESMS (C₂₂H₄₀N₆O₁); calcd. 404.6;obsd. 405.4 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.05-7.0 (d,2H), 6.98-6.9 (t, 1H), 3.98-3.96 (d, 2H), 3.8-3.70 (m, 1H), 3.55-3.46(br s, 4H), 3.44-4.34 (br s, 4H), 3.32-3.20 (m, 4H), 3.18-3.05 (m, 4H),2.29 (s, 6H), 2.30-2.15 (m, 2H), 2.05-1.95 (m, 2H).

Compound 3 was prepared from 1,8-diamino-3,6-dioxaoctane in an analogousmanner as described above. ESMS (C₁₈H₃₂N₄O₃); calcd352.5; obsd. 353.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.04-7.0 (d, 2H), 6.98-6.9(t, 1H), 3.99-3.96 (d, 2H), 3.85-3.83 (t, 2H), 3.82-3.75 (m, 1H),3.72-3.68 (m, 4H), 3.65-3.51 (t, 2H), 3.45-3.40 (t, 2H), 3.47-3.45 (t,2H), 2.29 (s, 6H), 1.53-1.51 (d, 3H).

Compound 4 was prepared from 1,5-diaminopeptane in an analogous manneras described above. ESMS (C₁₇H₃₀N₄O₁); calcd. 306.8; obsd. 307.3 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.02-7.0 (d, 2H), 6.97-6.91 (t, 1H),4.0-3.9 (m, 2H), 3.79-3.65 (br m, 1H), 3.21-3.15 (m, 4H), 2.28 (s, 6H),1.84-1.78 (br m, 2H), 1.7-1.6 (quin, 2H), 1.58-1.5 (m, 5H). Retentiontime (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=2.2 min.

Compound 5 was prepared from 1,13-diamino-4,7,10-trioxatridecane in ananalogous manner as described above. ESMS (C₂₂H₄₀N₄O₄); calcd. 424.6;obsd. 425.2 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.03-7.0 (d,2H), 6.98-6.91 (t, 1H), 4.0-3.9 (m, 2H), 3.80-3.78 (m, 1H), 3.75-3.65(t, 2H), 3.62-3.58 (m, 4H), 3.56-3.5 (m, 6H), 3.40-3.21 (m, 4H), 2.30(s, 6H), 2.05-2.01 (quin, 2H), 1.83-1.78 (quin, 2H), 1.50-1.48 (d, 3H).Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=2.51 min.

Compound 6 was prepared from N,N-bis(2-aminoethyl)methylamine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=0.38. ESMS (C₁₇H₃₁N₅O); calcd. 321.46; obsd.322.2 [M+H]⁺.

Compound 7 was prepared from 1,7-diamino-4-oxaheptane in an analogousmanner as described above. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=0.59. ESMS (C₁₈H₃₂N₄O₂); calcd. 336.47; obsd. 337.5[M+H]⁺.

Compound 8 was prepared from piperazine in an analogous manner asdescribed above. ESMS (C₁₆H₂₆N₄O₁); calcd. 290.4; obsd. 291.1 [M+H]⁺.Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=1.76 min.

Compound 9 was prepared from α,α′-diamino-m-xylene in an analogousmanner as described above. ESMS (C₂₀H₂₈N₄O₁); calcd. 340.5; obsd. 341.2[M+H]⁺. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=2.15min.

Compound 10 was prepared from 1,5-diamino-3-mercaptopentane m ananalogous manner as described above. ESMS (C₁₆H₂₈N₄O₁S₁); calcd. 324.5;obsd. 325.3 [M+H]⁺. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=2.05 min.

Compound 11 was prepared from α,α′-di-amino-p-xylene in an analogousmanner as described above. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=1.07. ESMS (C₂₀H₂₈N₄O); calcd. 340.46; obsd. 341.4[M+H]⁺.

Compound 12 was prepared from 1,4-diaminobutane in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=1.70. ESMS (C₁₆H₂₈N₄O); calcd. 292.42; obsd. 293.1 [M+H]⁺.

Compound 13 was prepared from 1,5-diamino-3-oxapentane in an analogousmanner as described above. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=1.68. ESMS (C₁₆H₂₈N₄O₂); calcd. 308.48; obsd. 309.3[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.04-7.02 (d, 2H), 6.97-6.92(t, 1H), 3.99-3.97 (d, 2H), 3.87-3.84 (t, 4H), 3.71-3.69 (t, 4H),3.47-3.40 (p, 1H), 2.30 (s, 6H), 1.53-1.51 (d, 3H).

Compound 14 was prepared from 1,2-diaminoethane in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=1.54. ESMS (C₁₄H₂₄N₄O); calcd. 264.37; obsd. 265.2 [M+H]⁺.

EXAMPLE 39 Synthesis of dimers of 5-arylpyrimidine and Mexiletine viathe Following Scheme

General procedure for synthesis of compound 4 of Table H: L═(CH₂)₅: ToTable G, compound 4 (0.488 g, 0.912 mmole) in a vial was added NaOMe(100 mg/mL; 1.0 mL, 1.85 mmole) and compound 12 (500 mg/mL MeOH; 0.5 mL,0.912 mmole). The reaction vessel was well sealed with a screw cap, andshaken at 80° C. for 12 h. The mixture was then concentrated to dryness,and the dark brown residue was dissolved in 5 mL of 50% aqueousacetonitrile (with 5% TFA). After filtration, the crude product waspurified by preparative reversed phase HPLC: 20 to 60% aqueousacetonitrile (0.1% TFA) over 50 min (linear gradient); 40 mL/min; 254nm. The desired product was obtained as white solid (80 mg). ESMS(C₂₇H₃₄N₅OFCl₂); calcd. 534.5; obsd. 534.2 [M]⁺. ¹H-NMR (CD₃OD, 299.96MHz): δ (ppm) 7.78-7.7 (dd, 1H), 7.5-7.4 (t, 1H), 7.38-7.3 (dd, 1H),7.05-7.01 (d, 2H), 6.97-6.9 (t, 1H),4.96 & 4.81 (two s, 2H), 4.18-4.05(t, 2H), 4.01-3.9 (m, 2H), 3.8-3.7 (m, 1H), 3.25-3.2 (t, 2H), 2.29 (s,6H), 1.95-1.79 (br m, 4H), 1.65-1.58 (br m, 2H), 1.52-1.50 (d, 3H).Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.48 min.

Table H lists linkers 1-14 from dimer H

TABLE H No. Linker 1 —(CH₂)₆— 2

3 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 4 —(CH₂)₅— 5—(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— 6 —(CH₂)₂—N(CH₃)—(CH₂)₂— 7—(CH₂)₃—O—(CH₂)₃— 8

9 —CH₂—Z—CH₂— where Z = 1,3-phenyl 10 —(CH₂)₂—S—(CH₂)₂— 11 —CH₂—Z—CH₂—where Z = 1,4-phenyl 12 —(CH₂)₄— 13 —(CH₂)₂—O—(CH₂)₂— 14 —(CH₂)₂—

Compound 1 was prepared from compound 1 of Table G in an analogous wayas described above. ESMS (C₂₈H₃₆N₅OFCl₂); calcd. 548.5; obsd. 548.3[M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (dd, 1H), 7.56-7.42(t, 1H), 7.33-7.3 (dd, 1H), 7.02-7.0 (d, 2H), 6.98-6.9 (dd, 1H), 4.96 &4.81 (two s, 2H), 4.18-4.1 (t, 2H), 4.02-3.9 (m, 2H), 3.78-3.7(m, 1H),3.21-3.17 (t, 2H), 2.29 (s, 6H), 1.9-1.75 (br m, 4H), 1.6-1.5 (m, 7H).Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.14 min.

Compound 2 was prepared from compound 2 of Table G in an analogous wayas described above. ESMS (C₃₂H₄₄N₇OFCl₂); calcd. 632.7; obsd. 632.4[M]⁺. ¹H-NMR (CD₃OD, 299.96. MHz): δ (ppm) 7.75-7.7 (dd, 1H), 7.5-7.4(t, 1H), 7.38-7.35 (dd, 1H), 7.05-7.0 (d, 2H), 6.98-6.9 (dd, 1H), 4.98 &4.82 (two s, 2H), 4.2-4.15 (t, 2H), 4.0-3.97 (m, 2H), 3.8-3.75 (m, 1H),3.35-3.2 (m, 6H), 3.19-3.1 (m, 6H), 2.95-2.88 (t, 2H), 2.30 (s, 6H),2.2-2.18 (m, 4H), 1.52-1.49 (d, 3H). Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=3.13 min.

Compound 3 was prepared from compound 3 Table G in an analogous way asdescribed above. ESMS (C₂₈H₃₆N₅O₃FCl); calcd.580.5; obsd. 580.4 [M]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (dd, 1H), 7.45-7.4 (t,1H), 7.35-7.3 (dd, 1H), 7.05-7.0 (d, 2H), 6.98-6.95 (dd, 1H), 4.96 &4.81 (two s, 2H), 4.4 (br m, 2H), 4.40-3.95 (m, 3H), 3.8-3.7 (m, 8H),3.42-3.4 (t, 2H), 2.29 (s, 6H), 1.52-1.49 (d, 3H). Retention time (anal.HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.40 min.

Compound 5 was prepared from compound 5 of Table G in an analogous wayas described above. ESMS (C₃₂H₄₄N₅O₄FCl₂); calcd. 652.6; obsd. 652.5[M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.71-7.7 (dd, 1H), 7.48-7.41(t, 1H), 7.35-7.32 (dd, 1H), 7.02-7.0 (d, 2H), 6.98-6.9 (dd, 1H₁), 4.97& 4.82 (two 4.2-4.14 (t, 2H), 4.0-3.9 (m, 3H), 3.8-3.5 (m, 12H),3.2-3.15 (m, 2H), 6H), 2.18-2.01 (m, 4H), 1.50-1.47 (d, 3H). Retentiontime (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.46 min.

Compound 6 was synthesized from compound 6 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=1.48. ESMS (C₂₇H₃₅Cl₂FN₆O); calcd. 549.52; obsd. 550.4[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.70 (d, 1H), 7.46-7.42(t, 1H), 7.32-7.30 (d, 1H), 7.05-7.02 (d, 2H), 6.97-6.92 (t, 1H), 4.98(s, 1H), 4.82 (s, 1H), 4.33-4.30 (t, 2H), 3.99-3.96 (m, 2H), 3.78-3.74(p, 1H), 3.04-3.01 (t, 4H), 2.97-2.92 (t, 2H), 2.47 (s, 3H), 2.29 (s,6H), 1.52-1.50 (d, 3H).

Compound 7 was synthesized from compound 7 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=1.56. ESMS (C₂₈H₃₆Cl₂FN₅O₂); calcd. 564.53; obsd. 565.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.71 (d, 1H), 7.48-7.43(t, 1H), 7.32-7.29 (d, 1H), 7.04-7.01 (d, 2H), 6.97-6.92 (t, 1),5.16-5.07 (m, 1H), 5.01-4.92 (m, 1H), 4.03-3.90 (m, 2H), 3.82-3.76 (p,1H), 3.70-3.67 (t, 2H), 3.62-3.58 (t, 2H), 3.55-3.50 (t, 2H), 2.30 (s,6H), 2.12-2.03 (quin, 2H), 1.95-1.86 (quin, 2H), 1.50-1.48 (d, 3H).

Compound 8 was prepared from compound 8 of Table G in an analogous wayas described above. ESMS (C₂₆H₃₀N₅OFCl₂); calcd. 518.5; obsd. 518.2[M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.64 (dd, 1H), 7.42 (t, 1H),7.27 (dd, 1H), 7.06-7.0 (dd, 2H), 6.99-6.92 (dd, 1H), 5.03 & 4.87 (twom, 2H), 4.18-4.10 (m, 3H), 4.0-3.93 (m, 2H), 3.62 (br s, 6H), 2.31 (s,6H), 1.60-1.57 (d, 3H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.94 min.

Compound 9 was prepared from compound 9 of Table G in an analogous wayas described above. ESMS (C₃₀H₃₂N₅OFCl₂); calcd. 568.5; obsd. 568.3[M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.7-7.65 (dd, 1H), 7.62-7.56(m, 2H), 7.42-7.39 (m, 2H), 7.38-7.3 (m, 2H), 7.02-7.0 (d, 2H), 6.98-6.9(dd, 1H), 5.54 (s, 2H), 5.03 & 4.87 (two s, 2H), 4.44 (s, 2H), 4.03-4.01(d, 2H), 3.8-3.7 (br m, 1H), 2.30 (s, 6H), 1.59-1.56 (d, 3H). Retentiontime (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.48 min.

Compound 10 was prepared from compound 10 of Table G in an analogous wayas described above. ESMS (C₂₆H₃₂N₅OFCl₂S); calcd.552.5; obsd. 552.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.68 (dd, 1H), 7.5-7.4(t, 1H), 7.35-7.3 (dd, 1H), 7.02-7.0 (d, 2H), 6.98-6.95 (dd, 1H), 4.97 &4.82 (two s, 2H), 4.46 (t, 2H), 3.99-3.97 (d, 2H), 3.81-3.78 (m, 1H),3.5-3.42 (t, 2H), 3.20-3.1 (t, 2H), 3.08-3.02 (t, 2H), 2.29 (s, 6H),1.53-1.51 (d, 3H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=3.46 min.

Compound 11 was synthesized from compound 11 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.11. ESMS (C₃₀H₃₂Cl₂FN₅O); calcd. 568.52; obsd. 569.3[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.73-7.72 (d, 1H), 7.68-7.66(d, 1H), 7.49-7.44 (t, 1H), 7.37-7.34 (d, 2H), 7.38-7.36 (d, 2H),7.04-7.02 (d, 2H), 6.97-6.92 (t, 1H), 5.53 (s, 2H), 5.04 (s, 1H), 4.88(s, 1H), 4.44 (s, 1H), 4.02-4.01 (d, 2H), 3.75-3.73 (m, 1H), 2.29 (s,6H), 1.57-1.55 (d, 3H).

Compound 12 was synthesized from compound 12 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.06. ESMS (C₂₆H₃₂Cl₂FN₅O); calcd. 526.48; obsd. 521.3[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.70 (d, 1H), 7.48-7.42(t, 1H), 7.33-7.31 (d, 1H), 7.05-7.02 (d, 2H), 6.97-6.92 (t, 1H), 4.978(s, 1H), 4.81 (s, 1H), 4.18 (m, 2H), 4.01-3.92 (m, 2H), 3.78-3.70 (m,1H), 3.26-3.22 (m, 2H), 2.97 (s, 6H), 1.94 (m, 4H), 1.53-1.51 (d, 3H).

Compound 13 was synthesized from compound 13 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.12. ESMS (C₂₆H₃₂Cl₂FN₅O₂); calcd. 536.48; obsd. 537.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.69 (d, 1H), 7.46-7.40(t, 1H), 7.31-7.29 (d, 1H), 7.04-7.02 (d, 2H), 6.97-6.92 (t, 1H), 4.96(s, 1H), 4.80 (s, 1H), 4.45-4.42 (t, 2H), 3.99-3.96 (t, 2H), 3.99-3.97(d, 2H), 3.92-3.89 (t, 2H), 3.85-3.77 (m, 1H), 3.46-3.43 (t, 2H), 2.29(s, 6H), 1.52-1.50 (d, 3H).

Compound 14 was synthesized from compound 14 of Table G in an analogousway as described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.30. ESMS (C₂₇H₃₅Cl₂FN₆O); calcd. 492.42; obsd. 493.2[M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.72-7.70 (d, 1H), 7.47-7.42(t, 1H), 7.32-7:30 (d, 1H), 7.03-7.01 (d, 2H), 6.96-6.91 (t, 1H), 4.99(s, 1H), 4.83 (s, 1H), 4.61-4.56 (t, 2H), 4.00-3.93 (m, 2H), 3.80-3.76(m, 1H), 3.69-3.65 (t, 2H), 2.29 (s, 6H), 1.51-1.49 (d, 3H).

EXAMPLE 40 Synthesis of Dimers of 5-arylpyrimidine and Mexiletine viathe Following Scheme

reagents and conditions: i)1-H-pyrazole-1-[N,N′-bis(tert-butoxycarbonyl)]-carboxamidine, MeCN, 80°C.; ii) TFA, CH₂Cl₂, rt

General procedure for the synthesis of the above compound 67A: To apyrazole-1-N,N′-bis(tert-butoxycarbonyl)carboxamidine (2.0 g, 6.44mole). The mixture was stirred at 80° C. for 14 h. The reaction mixturewas concentrated in vacuo, and the oily residue was dissolved in ethylether (100 mL), followed by washing with brine solution. The organicphase was dried over MgSO₄, and evaporated to yield a pale yellow oilyresidue (2.8 g). %): R_(f)=0.74 in 10% MeOH/EtOAc/hexane (1/1). It wasdissolved in CH₂Cl₂ (25 mL), and then followed by slow addition of TFA(13 mL) while stirring the mixture. After stirring overnight at rt, thereaction mixture was concentrated in vacuo, yielding colorless oilyresidue. The crude product was dissolved in 50% aqueous acetonitrile,and purified by preparative reversed phase. HPLC: 10 to 40% aq MeCN over50 min (linear gradient); 40 mL/min; 214 nm. Compound 67 was obtained ascolorless oil (1.0 g; overall 45% to two steps). ESMS (C₁₂H₁₉N₃O);calcd. 221.3; obsd. 222.0 [M+H]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm)7.01-6.99 (d, 2H), 6.93-6.9 (dd, 1H), 4.1-3.98 (m, 1H), 3.9-3.85 (dd,1H), 3.7-3.65 (dd, 1H), 2.26 (s, 6H), 1.39-1.37 (d, 3H). Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=2.91 mm.

Compound 68A was prepared from N-methyl-mexiletine in an analogous wayas described above. ESMS (C₁₃H₂₁N₃O); calcd. 235.3; obsd. 236.1 [M+H]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.01-6.99 (d, 2H), 6.93-6.89 (dd,1H), 4.45-3.35 (m, 1H), 3.98-3.95 (dd, 1H), 3.81-3.78 (dd, 1H), 3.05 (s,3H), 2.25 (s, 6H), 1.34-1.31 (d, 3H). Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=2.91 min.

Synthesis compound 2 of Table C: To compound 67 (TFA salt; 0.459 g, 1.37mmole) in a glass vial was added NaOMe (100 mg/mL; 0.75 mL, 1.39 mmole)and compound 12 (500 mg/mL MeOH; 1 mL, 1.82 mmole). The reaction vesselwas sealed well with a screw cap, and shaken at 80° C. for 12 h. Themixture was then concentrated to dryness, and the dark brown residue wasdissolved in 5 mL of 50% aqueous acetonitrile (with 5% TFA). Afterfiltration, the crude product was purified by preparative reverse phaseHPLC: 20 to 60% aqueous acetonitrile (0.1% TFA) over 50 min (lineargradient); 40 mL/min; 254 nm. The desired product was obtained as whitesolid (80 mg). ESMS (C₂₂H₂₃N₄OFCl₂); calcd. 449.4; obsd. 449.3 [M]⁺.¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.75-7.71 (dd, 1H), 7.5-7.4 (dt,1H), 7.38-7.32 (dd, 1H), 7.01-6.98 (d, 2H), 6.95-6.88 (dd, 1H), 5.25-4.9(two dd, 2H), 4.6 (br m, 1H), 4.0-3.8 (m, 2H), 2.27 (s, 6H), 1.51-1.49(d, 3H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=4.41min.

Compound 3 of Table C was synthesized from compound 68 in an analogousmanner as described above. ESMS (C₂₃H₂₅N₄OFCl₂); calcd. 463.4; obsd.463.1 [M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.75-7.7 (dd, 1H),7.5-7.42 (dt, 1H), 7.38-7.35 (dd, 1H), 7.0-6.97 (d, 2H), 6.95-6.88 (dd,1H), 5.21-4.95 (m, 2H), 4.16-4.0 (dd, 2H), 3.9-3.81 (m, 1H), 3.3 (s,3H), 2.25 (s, 6H), 1.42-1.4 (d, 3H), Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=4.21 min.

EXAMPLE 41 Synthesis Compound 4 of Table C via the Following Scheme

reagents and conditions: i)1-H-pyrazole-1-[N,N′-bis(tert-butoxycarbonyl)]carboxamidine, MeCN, 80°C.; ii) TFA, CH₂Cl₂, rt

Synthesis of Compound 500: To a solution of compound 100 (TFA salt; 1.0g, 3.0 mmole) in MeOH (5 mL) was added NaOMe (0.5 M, 6.31 mL). Themixture was concentrated to dryness in vacuo. The oily residue wassolubilized in THF (15 mL), and followed by addition of1-H-pyrazole-1-[N,N′-bis(tert-butoxycarbonyl)carboxamidine] (0.979 g,3.15 mmole). After stirring at 80° C. overnight, the reaction mixturewas diluted with EtOAc (150 mL) and then washed with brine solution. Theorganic phase was dried over Na₂SO₄, and evaporated to yield colorlessoily residue. It was dissolved in CH₂Cl₂ (15 mL), and then followed byslow addition of TFA (10 mL). The mixture was stirred overnight at rt,and was concentrated in vacuo, yielding colorless oily residue. Thecrude product was dissolved in 50% aqueous acetonitrile, and purified bypreparative reversed phase HPLC: 10 to 40% aq. MeCN over 40 min (lineargradient); 40 mL/min; 214 nm. Compound 69 was obtained as colorless oil.ESMS (C₁₅H₂₃N₃O); calcd. 261.4; obsd. 262.2 [M+H]⁺. ¹H-NMR (CD₃OD,299.96 MHz): δ (ppm) 7.01-6.98 (d, 2H), 6.91-6.85 (dd, 1H), 4.1-4.04 (brd, 1H), 3.85-3.78 (br d, 1H), 3.71-3.65 (m, 2H), 3.21-3.1 (m, 2H),2.2-1.95 (m, 2H), 1.89-1.82 (m, 1H), 1.7-1.55 (m, 2H). Retention time(anal. HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.35 min.

Synthesis of Table C, compounds 4: To compound 500 (TFA salt; 0.13 g,0.346 mmole) in a glass vial was added NaOMe (100 mg/mL; 0.22 mL, 0.407mmole) and compound 12 (500 mg/mL MeOH; 0.38 mL, 0.693 mmole). Thereaction vessel was sealed well with a screw cap, and shaken at 80° C.for 12 h. The mixture was then concentrated in vacuo, yielding the darkbrown residue. It was dissolved in 3 mL of 50% aqueous acetonitrile(with 5% TFA), filtered, and purified by preparative reversed phaseHPLC: 20 to 60% aqueous acetonitrile (0.1% TFA) over 50 min (lineargradient); 40 mL/min; 254 nm. ESMS (C₂₅H₂₇N₄OFCl₂); calcd. 489.5; obsd.489.2 [M]⁺. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=4.5 min.

EXAMPLE 42 Synthesis of Compounds 71, 72, and 73 via the FollowingScheme

reagents and conditions: i) (EtO)₂CHCO₂Et, KO^(t)BU, EtOH, rt; ii) Et-I;iii) guanidine hydrochloride, NaOMe, MeOH, 80° C.; iv)1,2-dimethoxyethane, 3M HCl, 90° C., 5 h.

Compounds 71, 72, and 73 were synthesized generally in accordance withprocedures described in Nobbs, et al., WO 97/09317 and Miller, et al.,EP 0 372 934B1.

Compound 71: To a stirred solution of compound 11 (10 g, 53.6 mmole) in1,2-diethoxyethane (75 mL) was added ethyl 2,2-diethoxyacetate (14.4 mL,80.5 mmole) and then KO^(t)Bu (7.2 g, 64.2 mmole) in ice bath. Thereaction mixture was gradually warmed to rt over 10 min, and then heatedat 90° C. for 6 h. After cooling the mixture to rt, ethyl iodide (8.6mL, 0.108 mole) was added, and followed by heating at 65° C. overnight.Reaction mixture was concentrated to brown solid, which was thenpartitioned between EtOAc (300 mL) and brine (200 mL). After shaking ina separatory funnel, organic phase was collected, washed with brinesolution, and dried over Na₂SO₄. Evaporation of the organic phaseafforded compound 71 as dark orange oil. ESMS (C₁₆H₁₉N₁O₃Cl₂); calcd.344.2; obsd. 345.1 [M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.42-7.4(dd, 1H), 7.31-7.28 (dd, 1H), 7.18-7.14 (t, 1H), 5.34 (s, 1H), 3.8-3.6(m, 6H), 1.33-1.19 (m, 9H).

Compound 72: To EtOH (80 mL) cooled in ice bath was added in smallportions NaOEt (6 g, 88.2 mmole) under stream of nitrogen, and thenfollowed by addition of guanidine hydrochloride (8.3 g, 86.9 mmole).After stirring at 0° C. for 10 min, compound 71 (15 g, 43.5 mmole) inEtOH (70 mL) was added to it. The mixture was warmed to rt over 2 h, andheated at 65° C. overnight. After concentration to brown oily residue,it was treated with cold water (200 mL) to yield brown precipitate. Thesupernatant was decanted, and the oily residue was washed with water(200 mL). The residue was dissolved in EtOAc (500 mL), washed with brinesolution, and dried over NaSO₄. Evaporation of the organic phaseafforded crude compound 7 as dark orange oil, which solidified slowlyover several days. The solid material was suspended in i-PrOH/hexane(1/1), and collected on Büchner funnel. This product was used in nextstep without further purification. ESMS (C₁₅H₁₈N₄O₂Cl₂); calcd. 357.6;obsd. 357.1 [M]⁺. ¹H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.61-7.58 (dd,1H), 7.38-7.35 (t, 1H), 7.23-7.21 (d, 1H), 4.78 (s, 1H), 3.78-3.35 (m,4H), 1.25-1.02 (m, 6H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=3.3 min.

Compound 73: To a solution of compound 72 (3 g, 8.39 mmole) in1,2-dimethoxymethane (70 mL) was added 70 mL of 3 M HCl, and the mixturewas heated at 90° C. for 5 h. The reaction mixture was concentrated, anddissolved in 50% aqueous acetonitrile prior to purification by usingpreparative reversed phase HPLC: 10 to 60% MeCN over 50 min; 40 ml/min;254 nm. Compound 73 (TFA salt) was obtained as pale beige solid. ESMS(C₁₁H₁₀N₄O₂Cl₂); calcd. 301.3; obsd. 301.4 [M]⁺. ¹H-NMR (CD₃OD, 299.96MHz): δ (ppm) 7.71-7.35 (dd, 1H), 7.48-7.41 (t, 1H), 7.35-7.29 (m, 1H),5.09 & 5.03 (two s, 1H). Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=0.90 min.

EXAMPLE 43 Synthesis of Compound 1 of Table I and Compound 5 of Table Cvia the Following Scheme

reagents and condition: i) RNH₂, EtOH, 70° C.; then NaCNBH₃, rt

Table C, Compound 5:

TABLE I

No. Linker R R′ R^(5a) R^(6a) R^(5b) R^(6b) 1 —(CH₂)₂—O— H H Cl Cl Cl Cl(CH₂)₂—

General procedure synthesis compound 1 of Table I: To a solution ofmexiletine (28 mg, 0.16 mmole) in EtOH (1 mL) was added compound 73 (TFAsalt; 41.5 mg, 0.1 mmole). The reaction mixture was stirred at70° C. for6 h, and cooled to rt prior to addition of NaCNBH₃ (13 mg, 0.21 mmole).After stirring for 2 h at rt, and concentration in vacuo, the crudeproduct was dissolved in aqueous acetonitrile and purified by reversedphase HPLC: 10 to 50% MeCN over 50 min; 10 mL/min; 254 nm. ESMS(C₂₂H₂₅N₅OCl₂); calcd. 446.4; obsd. 460.0 [M]⁺. Retention time (anal.HPLC: 10 to 70% MeCN/H₂O over 6 min)=3.20 min. ¹H-NMR (CD₃OD, 299.96MHz): δ (ppm) 7.69-7.66 (d, 1H), 7.46-7.35 (m, 1H), 7.34-7.32 (d, 1H),7.01-7.69 (d, 2H), 6.95-6.90 (t, 1H), 4.03-4.01 (d, 1H), 3.96-3.95 (d,1H), 3.89-3.88 (d, 2H), 3.75-3.69 (p, 1H), 2.17 (s, 6H), 1.42-1.40 (d,3H).

Compound 5 of Table C was prepared fromN-[1-(5-guanidino-3-oxa)pentyl]mexiletine in an analogous manner asdescribed above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over 6min)=3.21. ESMS (C₂₆H₃₄Cl₂N₆O₂); calcd. 533.49; obsd. 534.2 [M+H]⁺.

EXAMPLE 44 Synthesis Compounds 1 to 5 of Table J via the FollowingScheme

reagents condition: i) H₂N—L—NH₂, EtOH, 70° C.; then NaCNBH₃, rt

TABLE J No. Linker 1 —(CH₂)₃— 2 —(CH₂)₃—O—(CH₂)₃— 3 —CH₂—C(CH₃)₂—CH₂— 4—CH₂-Z-CH₂— where Z = 1,4-cyclohexyl 5 —CH₂-Z-CH₂— where Z = 1,3-phenyl6 —(CH₂)₄— 7 —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— 8 —CH₂-Z-CH₂— where Z =trans-1,4-cyclohexyl

Compound 1 of Table J: To a solution of 1,3-diaminopropane (3.4 mg, 0.05mmole) in EtOH (1 mL) was added compound 73 (TFA salt; 41.5 mg, 0:1mmole). The reaction mixture was stirred at 70° C. for 6 h, and cooledto rt prior to addition of NaCNBH₃ (13 mg, 0.21 mmole). After stirringfor 2 h at rt, and, concentration in vacuo, the crude product wasdissolved in aqueous acetonitrile and purified by reversed phase HPLC.ESMS (C₂₅H₂₆N₁₀Cl₄); calcd. 608.4; obsd. 609.0 [M+H]⁺.

Compound 2 was prepared from 1,7-dimamino-4-oxaheptnane in a analogousmanner as described above. ESMS (C₂₈H₃₂N₁₀OCl₄); calcd. 666.4; obsd.667.1 [M+H]⁺.

Compound 3 was prepared from 2,2-dimethyl-1,3-diaminopropane in ananalogous manner as described above. ESMS (C₂₇H₃₀N₁₀Cl₄); calcd. 636.4;obsd. 637.4 [M+H]⁺.

Compound 4 was prepared from cyclohexane-1,4-dimethylamine in ananalogous manner as described above. ESMS (C₃₀H₃₄N₁₀Cl₄); calcd. 676.5;obsd. 677.1 [M+H]⁺.

Compound 5 was prepared from α,α′-dimamino-m-xylene in an analogousmanner as described above. ESMS (C₃₀H₂₈N₁₀Cl₄); calcd. 670.4; obsd. 671[M+H]⁺.

Compound 6 was prepared from 1,4-dimaminobutane in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=1.87. ESMS (C₂₆H₂₈Cl₄N₁₀); calcd. 622.38; obsd. 623.2 [M+H]⁺.

Compound 7 was prepared from trans-1,4-diaminocyclohexane in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=2.91. ESMS (C₂₈H₃₀Cl₄N₁₀); calcd. 648.42; obsd.649.0 [M+H]⁺.

Compound 8 was prepared from trans-1,4-diaminocyclohexane in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=2.91. ESMS (C₂₈H₃₂Cl₄N₁₀O₂); calcd. 682.43;obsd.683.2 [M+H]⁺.

EXAMPLE 45 Synthesis of Compound A1 via the Following Scheme

reagents and conditions: i) Ph₃P, DEAD, THF, ii) DCM, TFA

reagents and conditions: i) Dihalide, Diisopropylethylamine, DMP, 90C.

To a stirring, 0° C. solution of 2,6-dimethylphenol (12.2 g, 100 mmol),N-Boc-2-piperidineethanol (22.9 g, 100 mmol), and triphenylphosphene(29.6 g, 113 mmol) in 400 mL THF, was added dropwise over 30 min 19.7 mL(113 mmol) DEAD. The mixture was mixed slowly and allowed to warm toambient temperature and stirred for 12 hours. The mixture wasconcentrated in vacuo, and Hexane/DCM was added to precipitate outtriphenylphosphene oxide, which was filtered off. The filtrate wasconcentrated in vacuo, and the residue was flash chromatographed onsilica gel, the appropriate fractions were combined to give the desiredboc-amino ether. 10 g of this material was dissolved in 300 mL of DCM,cooled to 0° C., and 50 mL of TFA was added dropwise over the period of30 minutes. The mixture was allowed to warm to room temperature over 2hours, then was concentrated, then titrated with DCM to give the desiredamine as an off-white solid.

Compound A1

1H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.09-6.97 (d, 2H), 6.88-6.78 (dd,1H), 3.98-3.71 (m, 2H), 3.50-3.38 (d, 2H), 3.10-2.97 (t, 1H), 2.29-2.10(m, 7H), 2.05-1.85 (m, 3H), 1.75-1.49 (m, 3H). ESMS (C₁₅H₂₃NO): calcd.233.35, obsd. 234.0 [M+H]+.

Synthesis of Dimers of Compounds A1

A general Procedure (synthesis of Compound A2 which is a dimer of A1): Asolution of Compound A1 (69.5 mg of the TFA salt, 0.2 mmol) withdiisopropylethylamine (108 μl, 0.6 mmol) in 250 μL anhydrous DMF, wasadded to a solution of 1,3-diiodo-2propanol (31.8 mg, 0.1 mmol) in 250μL anhydrous DMF. The mixture was shaken for 20 h at 90° C., thenstripped of solvent under vacuum. The resulting tarry mixture wasdissolved in 1 mL of a 1:1 mixture of acetonitrile and water, with 0.1%trifluoroacetic acid. This mixture was separated by preparative HPLC.

Compounds A2

Retention Time (anal. HPLC: 5-55% MeCN/H₂O over 5 min)=4.90 min. ESMS(C₃₃H₅₀N₂O₃): calcd. 522.78, obsd. 523.6 [M+H]+.

Synthesis of Compounds A3-A12 Which Have Different Linkers L IdentifiedAbove

Compound A3 was prepared in an analogous manner frombis-iodoethoxyethane

Retention Time (anal. HPLC: 5-55% MeCN/H₂O over 5 min)=4.85 min. ESMS(C₃₆H₅₆N₂O₄): calcd. 580.85, obsd. 581.6 [M+H]+.

Compound A4 was prepared in an analogous manner from 1,7-dibromoheptane

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.99 min. ESMS(C₃₆H₅₆N₂O₄): calcd. 562.88, obsd. 563.6 [M+H]+.

Compound A5 was prepared in analogous manner from 1,8-dibromooctane

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=4.06 min. ESMS(C₃₆H₅₆N₂O₄): calcd. 576.91, obsd. 577.6 [M+H]+.

Compound A6 was prepared in an analogous manner from 1,9-dibromononane

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=4.14 min. ESMS(C₃₆H₅₆N₂O₄): calcd. 590.93, obsd. 591.6 [M+H]+.

Compound A7 was prepared in an analogous manner from3-chloro-2-chloromethyl-1-propene, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.97 min. ESMS(C34H50N2O2): calcd. 518.78, obsd. 519.6 [M+H]+.

Compound A8 was prepared in an analogous manner from1,3-dibromo-m-xylene.

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.96 min. ESMS(C₃₈H₅₂N₂O₂): calcd. 568.84, obsd. 569.4 [M+H]+.

Compound A9 was prepared in an analogous manner from1,3-dibromo-p-xylene.

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.90 min. ESMS(C₃₈H₅₂N₂O₂): calcd. 568.84, obsd. 569.4 [M+H]+.

Compound A10 was prepared in an analogous manner frombis(4-chlorobutyl)ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.94 min. ESMS(C₃₈H₆₀N₂O₃): calcd. 592.91, obsd. 593.6 [M+H]+.

Compound A11 was prepared in an analogous manner frombis[2-(2-chloroethoxy)ethyl]ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 5-90% MeCN/H₂O over 5 min)=3.85 min. ESMS(C₃₈H₆₀N₂O₃): cald. 624.90, obsd. 625.6 [M+H]+.

Compound A12 was prepared in an analogous manner fromN,N′-bis(2-chloroethyl)oxamide, using a catalytic amount of NaI.

Retention Time (anal. HPILC: 5-90% MeCN/H₂O over 5 min)=3.66 min. ESMS(C₃₆H₅₄N₄O₄): calcd. 606.84, obsd. 607.6 [M+H]+.

EXAMPLE 46 Synthesis of Compound A13 via the Following Scheme

reagents and conditions: i) Ph₃P, DEAD, THF, ii) DCM, TFA

reagents and conditions: i) Dihalide, Diisopropylethylamine, DMF, 90C.

To a stirring, 0° C. solution of 2,6-dimethylphenol (11.86 g, 97.1mmol), N-Boc-3-pyrrolidinol (18.18 g, 97.1 mmol), and triphenylphosphene(28.85 g, 110 mmol) in 400 mL THF, was added dropwise over 30 min 15.4mL (110 mmol) DEAD. The mixture was mixed slowly and allowed to warm toambient temperature and stirred for 12 hours. The mixture wasconcentrated in vacuo, and Hexane/DCM was added to precipitate outtriphenylphosphene oxide, which was filtered off. The filtrate wasconcentrated in vacuo, and the residue was flash chromatographed onsilica gel, the appropriate fractions were combined to give the desiredboc-amino ether. 11.14 g of this material was dissolved in 300 mL ofDCM, cooled to 0° C., and 5 mL of TFA was added dropwise over the periodof 30 minutes. The mix was allowed to warm to room temperature over 2hours, then was concentrated, then titrated with DCM to give the desiredamine as red oil.

Compound A13

1H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.06-7.00 (d, 2H), 6.98-6.90 (dd,1H), 4.82-4.76 (m, 1H), 3.64-3.43 (m, 4H), 2.37-2.25 (m, 7H), 2.20-2.08(m, 1H). ESMS (C₁₂H₁₇NO): calcd. 191.27, obsd. 192.0 [M+H]+.

Synthesis of Dimers of Compound A13

A general procedure (synthesis of Compound A14 which is a dimer of A13):A solution of Compound A13 (61.1 mg of the TFA salt, 0.2 mmol) withdiisopropylethylamine (108 μl, 0.6 mmol) in 250 μL anhydrous DMF, wasadded to a solution of 1,3-diiodo-2propanol (31.8 mg, 0.1 mmol) in 250μL anhydrous DMF. The mixture was shaken for 20 h at 90° C., thenstripped of solvent under vacuum. The resulting tarry mixture wasdissolved in 1 mL of a 1:1 mixture of acetonitrile and water, with 0.1%trifluoroacetic acid. This mixture was separated by preparative HPLC.

Compound A14

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.54 min. ESMS(C₂₇H₃₈N₂O₃): calcd. 438.61, obsd. 439.5 [M+H]+.

Synthesis of Compounds A15-A22 Which Have Different Linkers L IdentifiedAbove

Compound A15 was prepared in an analogous manner from(−)-trans-4,5-bis(iodomethyl)-2,2-dimethyl-1,3-dioxolane

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=4.12 min. ESMS(C₃₁H₄₄N₂O₄): calcd. 508.71, obsd. 509.4 [M+H]+.

Compound A16 was prepared in an analogous manner frombis-iodoethoxyethane

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=3.84 min. ESMS(C₃₀H₄₄N₂O₄): calcd. 496.69, obsd. 497.4 [M+H]+.

Compound A17 was prepared in an analogous manner from1,3-dibromo-p-xylene.

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=4.01 min. ESMS(C₃₂H₄₀N₂O₂): calcd. 484.68, obsd. 485.4 [M+H]+.

Compound A18 was prepared in an analogous manner frombis(4-chlorobutyl)ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=4.06 min. ESMS(C₃₂H₄₈N₂O₃): calcd. 508.74, obsd. 509.4 [M+H]+.

Compound A19 was prepared in an analogous manner frombis[2-(2-chloroethoxy)ethyl]ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-90% MeCN/H₂O over 5 min)=3.27 min. ESMS(C₃₂H₄₈N₂O₅): calcd. 540.74, obsd. 541.4 [M+H]+.

Compound A20 was prepared in an analogous manner fromN,N′-bis(2-chloroethyl)oxamide, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=3.77 min. ESMS(C₃₀H₄₂N₄O₄): calcd. 522.69, obsd. 523.4 [M+H]+.

Compound A21 was prepared in an analogous manner from4,4-bis(chloromethyl)1,1-biphenyl, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-60% MeCN/H₂O over 5 min)=4.59 min. ESMS(C₃₈H₄₄N₂O₂): calcd. 560.78, obsd. 561.4 [M+]+.

Compound A22 was prepared in an analogous manner from 1,3-diiodopropane

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.54 min. ESMS(C₂₇H₃₈N₂O₂): calcd. 422.61, obsd. 423.4 [M+H]+.

EXAMPLE 47 Synthesis of Compound A23 via the Following Scheme

reagent and conditions: i) Ph₃P, DEAD, TBF, ii) DCM, TFA

reagents and conditions: i) Dihalide, Diisopropylethylamine, DMF, 90C.

To a stirring, 0° C. solution of 2,6-dimethylphenol (3.15 g, 25.8 mmol)N-Boc-3-hydroxypiperidine (5.2 g, 25.8 mmol), and triphenylphosphene(7.64 g, 29.15 mmol) in 100 mL TBF, was added dropwise over 30 min 4.08mL (29.15 mmol) DEAD; The mixture was mixed slowly and allowed to warmto ambient temperature and stirred for 12 hours. The mixture wasconcentrated in vacuo, and Hexane/DCM was added to precipitate outtriphenylphosphene oxide, which was filtered off. The filtrate wasconcentrated in vacuo, and the residue was flash chromatographed onsilica gel, the appropriate fractions were combined to give the desiredboc-amino ether. 1.8 g of this material was dissolved in 300 mL of DCM,cooled to 0° C., and 50 mL of TFA was added dropwise over the period of30 minutes. The mixture was allowed to warm to room temperature over 2hours, then was concentrated, then titrated with DCM to give the desiredamine as a clear oil.

Compound A23

1H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.06-7.00 (d, 2H), 6.96-6.88 (dd,1H), 4.18-4.10 (m, 1H), 3.47-3.32 (m, 2H), 3.22-3.10 (m, 2H), 2.32-2.27(s, 6H), 2.27-2.14 (m, 1H), 2.01-1.70 (m, 3H). ESMS (C₁₃H₁₉NO): calcd.205.30, obsd. 206.1 [M+H]+.

Synthesis of Dimers of Compound A23

A general procedure (synthesis of Compound A24 which is a dimer of A23):A solution of Compound A23 (63.8 mg of the TFA salt, 0.2 mmol) withdiisopropylethylamine (108 μl, 0.6 mmol) in 250 μL anhydrous DMF, wasadded to a solution of 1,3-diiodo-2propanol (31.8 mg, 0.1 mmol) in 250μL anhydrous DMF. The mixture was shaken for 20 h at 90° C., thenstripped of solvent under vacuum. The resulting tarry mixture wasdissolved in 1 mL of a 1:1 mixture of acetonitrile and water, with 0.1%trifluoroacetic acid. This mixture was separated by preparative HPLC.

Compound A24

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.92 min. ESMS(C₂₉H₄₂N₂O₃): calcd. 466.66, obsd. 467.4 [M+H]+.

Synthesis of Compounds A25-A34 Which Have Different Linkers L IdentifiedAbove

Compound A25 was prepared in an analogous manner frombisiodoethoxyethane

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.86 min. ESMS(C₃₂H₄₈N₂O₄): calcd. 524.74, obsd. 525.4 [M+H]+.

Compound A26 was prepared in an analogous manner from 1,3-dilodopropane

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.88 min. ESMS(C₉H₄₂N₂O₂): calcd. 450.66, obsd. 451.2 [M+H]+.

Compound A27 was prepared in an analogous manner from 1,6-diiodohexane

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.99 min. ESMS(C₃₂H₄N₂O₂): calcd. 492.74, obsd. 493.4 [M+H]+.

Compound A28 was prepared in an analogous manner from 2,6-bis(bromomethyl) pyridine

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.94 min. ESMS(C₅₃H₄₃N₃O₂): calcd. 513.72, obsd. 514.4 [M+H]+.

Compound A29 was prepared in an analogous manner from3-chloro-2-chloromethyl-1-propene, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.09 min. ESMS(C₃₀H₄₂N₂O₂): calcd. 462.67, obsd. 463.2 [M+H]+.

Compound A30 was prepared in an analogous manner from1,3-dibromo-m-xylene.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.04 min. ESMS(C₃₄H₄₄N₂O₂): calcd. 512.73, obsd. 513.4 [M+H]+.

Compound A31 was prepared in an analogous manner from1,3-dibromo-p-xylene.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.94 min. ESMS(C₃₄H₄₄N₂O₂): calcd. 512.73, obsd. 513.4 [M+H]+.

Compound A32 was prepared in an analogous manner from bis(4-chlorobutyl) ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=4.00 min. ESMS(C₃₄H₆₂N₂O₃): calcd. 536.80, obsd. 537.4 [M+H]+.

Compound A33 was prepared in an analogous manner from bis[2-(2-chloroethoxy) ethyl] ether, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.84 min. ESMS(C₃₄H₅₂N₂O₂): calcd. 568.80, obsd. 569.4 [M+H]+.

Compound A34 was prepared in an analogous manner from N,N′-bis(2-chloroethyl) oxamide, using a catalytic amount of NaI.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.77 min. ESMS(C₃₂H₄₆N₄O₄): calcd. 550.74, obsd. 551.4 [M+H]+.

EXAMPLE 48 Synthesis of Compound A35 via the Following Scheme

reagents and conditions: i) chloroacetone, K₂CO₃, KI, DMF,150C, ii)Toluene, bisiodoethioxyethane, 75C iii) NaBH₃CN, r.t.

To a stirring, 150° C. solution of 4-chloro-2,6-dimethylphenol (15.66 g,100 mmol), potassium carbonate (14.0 g, 100 mmol), and potassium iodide(2.0 g, catalytic amount) in 400 mL DMEF, was added dropwise over 30 min12.0 mL (150 mmol) chloroacetone. The mixture was continuously heated at150° C. and stirred for 12 hours. The mixture was filtered, and thenconcentrated in vacuo to give a tarry residue that was taken up in ethylacetate and water washed. This solution was concentrated and the residuediluted with hexanes, which caused a brown precipitate to form, whichwas filtered off. The filtrate was filtered through a pad of basicalumna to remove unreacted phenol. The filtrate was concentrated toprovide the desired ketone as a pale yellow solid.

Compound A35

1H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 7.03 (s, 2H), 4.49 (s, 2H), 2.23 (s,9H).

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.96 min.

Synthesis of Compound A36

To a solution of compound A35 (212.7 mg, 1.0 mmol) in 2 mL anhydroustoluene, was added 2,2′-(ethylenedioxy)bis(ethylamine) (73 μL, 0.5mmol), followed by 4A molecular sieves and 50 mg sodium sulfate. Themixture was shaken for 12 h at 75° C., then a solution of NaBH3CN (125.7mg, 2.0 mmol) in ethanol was added, and the mixture shaken for 2 hoursat 25° C. The mix was then quenched with water, concentrated underreduced pressure, then dissolved in 1 mL of a 1:1 mixture ofacetonitrile and water, with 0.1% trifluoroacetic acid. This mixture wasseparated by preparative HPLC.

Compound A36

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.00 min. ESMS(C₂₈H₄₂C₁N₂O₄): calcd. 541.55, obsd. 542.2 [M+H]+.

EXAMPLE 49 Synthesis of Compound A38 via the Following Scheme

reagents and conditions: i) Toluene, N-Me Mexiletine, 75C, ii) NaBH₃CN,r.t.

To a solution of compound 1 (178 mg, 1 mmol) in 2 mL anhydrous toluene,was added a solution of neutral N-methyl mexiletine (193.3 mg, 1 mmol)in 2 mL anhydrous toluene, followed by, 4A molecular sieves and 50 mgsodium sulfate. The mixture was shaken for 12 h at 75° C., then asolution of NaBH3CN (125.7 mg, 2.0 mmol) in ethanol was added, and themix shaken for 2 hours at 25° C. The mix was then quenched with water,concentrated under reduced pressure, then dissolved in 1 mL of a 1:1mixture of acetonitrile and water, with 0.1% trifluoroacetic acid. Thismixture was separated by preparative HPLC.

Compound A37

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.92 min. ESMS(C₂₃H₃₃NO₂): calcd. 355.52, obsd. 356.4 [M+H]+.

EXAMPLE 50 Synthesis of Compound A39 via the Following Scheme

reagents and conditions: i) Ph₃P, DEAD

reagents and conditions: i) ethanol, diamine, ii) NaBH₃CN, ii) TFA/DCM

To a stirring, 0° C. solution of 3,5-dimethyl-4-hydroxybenzaldehyde(1.5018 g, 10 mmol), N-Boc-3-piperidineethanol (2.153 g, 10 mmol), andtriphenylphosphene (2.964 g, 11.3 mmol) in 40 mL THF, was added dropwiseover 10 min 1.78 mL (11.3 mmol) DEAD the mixture was mixed slowly andallowed to warm to ambient temperature and stirred for 12 hours. Themixture was concentrated in vacuo, and Hexane/DCM was added toprecipitate out triphenylphosphene oxide, which was filtered off. Thefiltrate was concentrated in vacuo, and the residue was flashchromatographed on silica gel, the appropriate fractions combined togive the desired boc-amino ether.

Compound A39

1H-NMR (CD₃OD, 299.96 MHz): δ (ppm) 9.82 (s, 1H), 7.60 (s, 21),4.29-4.20 (dd, 1H), 3.90-3.98 (d, 1H), 3.0-2.78 (m, 2H), 2.35 (s, 6H),2.09-2.0 (m, 2H), 1.95-1.87 (m, 1H), 1.78-1.65 (m, 1H), 1.58-1.40 (m,10H). ESMS (C₂₀H₂₉NO₄): calcd. 347.45, obsd. 348.2 [M+H]+.

Synthesis of Dimers of Compound A39

A general procedure (synthesis of Compound A40 which is a dimer of A39):A solution of compound A39 (69.5 mg, 0.2 mmol) in 250 μL anhydrous EtOH,was added 1,8-diamino-3,6-dioxaoctane (14.8 mg, 0.1 mmol). The mixturewas shaken for 12 h at 23° C., then a solution of NaBH4 (15.2 mg, 0.4mmol) in ethanol was added, and the mix shaken for 2 hours at 25° C. Themix was then quenched with a solution of 5% trifluoroacetic acid inacetonitrile/water (1:1), concentrated under reduced pressure, thendissolved in 1 mL of a 10:1 mixture of dichloromethane/trifluoroaceticacid, and shaken for 12 hours. This was then concentrated under reducedpressure, then dissolved in 1 mL of a 1:1 mixture of acetonitrile andwater, with 0.1% trifluoroacetic acid. This mixture was separated bypreparative HPLC.

Compound A40

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.84 min. ESMS(C₃₆H₅₈N₄O₄): calcd. 610.88, obsd. 611.6 [M+H]+.

Synthesis of Compounds A41-A45 Which Have Different Linkers L IdentifiedAbove

Compound A41 was prepared in an analogous manner from neutralmexiletine.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=3.45 min. ESMS(C₂₆H₃₈N₂O₂): calcd. 410.6, obsd. 411.2 [M+H]+.

Compound A42 was prepared in an analogous manner fromN,N′-bis(3-aminopropyl) methylamine.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.27 min. ESMS(C₃₇H₆₁N₅O₂): calcd. 607.92, obsd. 608.4 [M+H]+.

Compound A4 was prepared m an analogous manner from1,3-cyclohexane-bis(methylamine).

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.73 min. ESMS(C₃₈H₆₀N₄O₂): calcd. 604.92, obsd. 605.6 [M+H]+.

Compound A44 was prepared in an analogous manner from 1,6-hexanediamine.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.63 min. ESMS(C₃₆H₅₈N₄O₂): calcd. 578.88, obsd. 579.6 [M+H]+.

Compound A45 was prepared in an analogous manner from p-xylenediamine.

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.66 min. ESMS(C₃₈H₅₄N₄O₂): calcd. 598.87, obsd. 599.4 [M+H]+.

EXAMPLE 51 Synthesis of Compound A46 via the Following Scheme

reagents and conditions: i) Ph₃P, DEAD ii) TFA/DCM

To a stirring, 0° C. solution of3,3′,5,5′-tetramethyl-[1,1′-biphenyl]-4,4′-diol (1.2116 g, 5.0 mmol),N-Boc-3-piperidineethanol (2.153 g, 10 mmol), and triphenylphosphene(2.964 g, 11.3 mmol) in 40 mL THF, was added dropwise over 10 min 1.78mL (11.3 mmol) DEAD. Mix slowly allowed to warm to ambient temperatureand stirred for 12 hours. The mixture was concentrated in vacuo, andHexane/DCM was added to precipitate out triphenylphosphene oxide, whichwas filtered off. The filtrate was concentrated in vacuo, and theresidue was flash chromatographed on silica gel, the appropriatefractions combined to give the desired boc-amino ether. This compound(3.0 g, 4.7 mmol) was dissolved in 100 mL Dichloromethane and cooled to0° C. in an ice bath. 10 mL of trifluoroacetic acid was added dropwisewith stirring over 20 minutes, and the mixture was slowly brought toroom temperature. The mixture was concentrated in vacuo and the excessTFA was removed by toluene azeotroping to give the product as a whitesolid.

Compound A46

Retention Time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.83 min. ESMS(C₂₈H₄₀N₂O₂): calcd. 436.64, obsd. 437.5 [M+H]+.

EXAMPLE 52 Heterodimer Starting Material (compounds B1-B8)

reagent and conditions: i) H₂N—Ln—NH₂, EtOH, molecular seives NaSO₄, 70°C.; then NaCNBH₃, rt

General procedure synthesis compound B1 via the above schemes. To asolution of 4,9-Dioxa-1,12-dodecanediamine (1.192 ml, 5.61 mmole) inEtOH (1.5 ml) was added compound SM1 (1.0 g, 5.61 mmole), molecularsieves (˜10 sieves), and Na₂SO₄ (20.0 mg). The reaction was stirred at70° C. for 6 h and then cooled to room temperature prior to addition ofNaCNBH₃ (529.1 mg, 8.42 mmole). After stirring for 2 h at rt, andconcentrating in vacuo, the crude product was dissolved in aqueousacetonitrile and purified by reversed phase HPLC: 10 to 50% MeCN over 60min; 40 ml/min; 214 nm. Retention time (anal. HPLC: 10 to 70% MeCN/H₂Oover 6 min)=2.15. ESMS (C₂₁H₃₈N₂O₃); calcd. 366.54; obsd. 367.2 [M+H]⁺.

Compound B2 was prepared from N-Methyl-2,2′diaminodiethylamine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=1.52. ESMS (C₁₆H₂₉N₃O); calcd. 279.42; obsd.280.2 [M+H]⁺.

Compound B3 was prepared from 2,2′-Tiobis(ethylamine) in an analogousmanner as described above. Retention time (anal. HPLC: 10 to+70%MeCN/H₂O over 6 min)=1.61. ESMS (C₁₅H₂₆N₂OS); calcd. 282.45; obsd. 283.2[N+H]⁺.

Compound B4 was prepared from m-Xylylenediamine in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=2.00. ESMS (C₁₉H₂₆N₂O); calcd. 298.42; obsd. 299.1 [M+H]⁺.

Compound B5 was prepared from 1,6-Diaminohexane in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=. ESMS (C₁₇H₃₀N₂O); calcd. 278.43; obsd. [M+H]⁺.

Compound B6 was prepared from Trans-1,4-cyclohexanediamine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=. ESMS (C₁₇H₂₈N₂O); calcd. 276.42; obsd.[M+H]⁺.

Compound B7 was prepared from 1,8-Diaminooctane in an analogous manneras described above. Retention time (anal. HPLC: 10 to 70% MeCN/H₂O over6 min)=. ESMS (C₁₉H₃₄N₂O); calcd. 306.49; obsd. [M+H]⁺.

Compound B8 was prepared from Bis (3-aminopropyl) ether in an analogousmanner as described above. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=. ESMS (C₁₇H₃₀N₂O₂); calcd. 294.43; obsd. [M+H]⁺.

EXAMPLE 53 Synthesis of Heterodimers 6-13

reagents and conditions: i) Mex—Ln—NH₂, EtOH, molecular seives, NA₂SO₄,70° C.; the NaCNBH₃, rt

General procedure synthesis compound 6 via the above scheme; To asolution of N-[1-(12-amino-4,9-dioxa)dodecyl]mexiletine (65.4 mg, 0.11mmole) in EtOH (1 ml) was added compound 73 (TFA salt; 40.0 mg, 0.096mmole), molecular sieves (6 sieves), and NaSO₄ (10.0 mg). The reactionwas stirred at 70° C. for 6 h and then cooled to room temperature priorto addition of NaCNBH₃ (78.6 mg, 0.125 mmole). After stirring for 2 h atrt, and concentrating in vacuo, the crude product was dissolved inaqueous acetonitrile and purified by reversed phase HPLC: 10 to 50% MeCNover 50 min; 10 ml/min; 254 nm. Retention time (anal. HPLC: 10 to 70%MeCN/H₂O over 6 min)=3.46. ESMS (C₃₂H₄₆Cl₂N₆O₃); calcd. 633.66; obsd.634.4 [M+H]⁺.

Compound 7 was prepared from N-[1-(2-amino-N-methyl)ethyl]mexiletine inan analogous manner as described above. Retention time (anal. HPLC: 10to 70% MeCN/H₂O over 6 min)=3.32. ESMS (C₂₇H₃₇Cl₂N₇O); calcd. 546.54;obsd. 547.3 [M+H]⁺.

Compound 8 was prepared from N-[1-(5-amino-3-thio)pentyl]mexiletine inan analogous manner as described above. Retention time (anal. HPLC: 10to 70% MeCN/H₂O over 6 min)=3.33. ESMS (C₂₆H₃₄Cl₂N₆OS); calcd. 549.57;obsd. 550.3 [M+H]⁺.

Compound 9 was prepared from N-[1(2-amino-m-xyleyl)methyl]mexiletine inan analogous manner as described above. Retention time (anal. HPLC: 10to 70% MeCN/H₂O over 6 min)=3.46. ESMS (C₃₀H₃₄Cl₂N₆O); calcd. 565.55;obsd. 566.2 [M+H]⁺.

Compound 10 was prepared from N-[1-(6-amino)hexyl]mexiletine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=3.38. ESMS (C₂₈H₃₈Cl₂N₆O); calcd. 545.56; obsd.546.1 [M+H]⁺.

Compound 11 was prepared from N-[1-(4-amino)cyclohexyl]mexiletine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=3.25. ESMS (C₂₈H₃₆Cl₂N₆O); calcd. 543.54; obsd.544.3 [M+H]⁺.

Compound 12 was prepared from N-[1-(8-amino)octyl]mexiletine in ananalogous manner as described above. Retention time (anal. HPLC: 10 to70% MeCN/H₂O over 6 min)=3.63. ESMS (C₃₀H₄₂Cl₂N₆O); calcd. 573.61; obsd.574.2 [M+H]⁺.

Compound 13 was prepared from N-[1-(7-amino-4-oxa)heptyl]mexiletine inan analogous manner as described above. Retention time (anal. HPLC: 10to 70% MeCN/H₂O over 6 min)=3.25. ESMS (C₂₈H₃₈Cl₂N₆O₂); calcd. 561.55;obsd. 562.1 [M+H]⁺.

Isolation and Purification of the Compounds

Isolation and purification of the compounds and intermediates describedherein can be effected, if desired, by any suitable separation orpurification such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography,thick-layer chromatography, preparative low or high-pressure liquidchromatography or a combination of these procedures. Characterization ispreferably by NMR and mass spectroscopy.

Utility and Testing

The multibinding compounds of this invention can be used to modulatesodium channels in various tissues including neurons, heart, and muscle.They will typically be used for the treatment of diseases and conditionsin mammals that involve or are mediated by Na⁺ channels, such aspathophysiological disorders, including hypertension, cardiacarrhythmogenesis, insulin-dependent diabetes, non-insulin dependentdiabetes mellitus, diabetic neuropathy, seizures, tachycardia, ischemiche disease, cardiac failure, angina, myocardial infarction, transplantrejection, autoimmune disease, sickle cell anemia, muscular dystrophy,gastrointestinal disease, mental disorder, sleep disorder, anxietydisorder, eating disorder, neurosis, alcoholism, inflammation,cerebrovascular ischemia, CNS diseases, epilepsy, Parkinson's disease,asthma, incontinence, urinary dysfunction, micturition disorder,irritable bowel syndrome, restenosis, subarachnoid hemorrhage,Alzheimers disease, drug dependence/addiction, schizophrenia,Huntington's chorea, tension-type headache, trigeminal neuralgia,cluster headache, migraine (acute and prophylaxis), depression, and theymediate the transmission of pain impulses by peripheral nerves, and thelike.

The multibinding compounds of this invention can be tested in well-knownand reliable assays and their activities are compared with those of thecorresponding unlinked (i.e., monovalent) ligands.

I. BIOASSAY OF THE EFFECT OF COMPOUNDS ON PAIN A. Acute Pain (theFormalin Test)

The formalin test is used as an animal model of acute injury. Asdescribed by Dubuisson and Dennis (1977, Pain 4:161), a standard dose offormalin is injected into the dorsal portion of the front paw of a rat.Bach rat is placed into a clear plastic cage for observation. Theanimals are observed and ratings, based on pain responses, are taken at30 and 60 minutes. Elevation, favoring, or excessive licking and bitingof the injected paw indicate a pain response. Analgesic response orprotection from compounds is indicated if both paws are resting on thefloor with no obvious favoring, excessive licking or biting of theinjected paw.

Determining the number of pain responses occurring per minutequantitates the analgesic effect of the test compounds. Theconcentration of test compound resulting in a 50% decrease inpain-responses/minute is the ED₅₀.

B. Chronic Pain

Modifications of the method of Hunter, J. C., et al., (1997, Eur. JPharmacol. 324:153) are used in the following models of chronic pain.

1. Constriction Injury

Adult male Spraque-Dawley rats weighing 150 g-180 g are placed two percage and allowed free access to food and water. The cages are housed intemperature and humidity controlled rooms and maintained on a 12-hlight/dark cycle.

Rats are anesthetized with sodium pentobarbital (70 mg/kg). Chronicconstriction injury is produced by exposing the common right sciaticnerve at mid-thigh level and proximal to the trifarcation of thesciatic. Four loose ligatures (4.0 chromic gut), with about 1-mmspacing, are tied around the nerve. The desired degree of constrictionretards but does not block circulation through the epineurialvasculatore. In every animal, an identical (sham) procedure is performedon the opposite side (left) with the exception that the sciatic nerve isnot ligated. All operations are completed by closing the muscle inlayers, applying wound clips to close the skin incision, and allowingthe animals to recover for a period of 5-7 days.

A cold water test is performed, after the recovery phase. The cold watertest is performed by placing each animal onto a metal stage submerged toa depth of 2.5 cm in ice-cold water (0° C.) contained within a squarePerspex chamber (21×21 cm). The animals respond by lifting the paw onthe ligated side out of the water. The latency to paw withdrawal(mean±standard error) is measured at 6.8±0.8 s (n=28). At no time doesany animal withdraw the paw on the sham side from the cold water. Ineach experiment, animals are first pre-screened twice with 20-mininterval between tests, in order to select for animals displaying clearsigns of cold allodynia, i.e. animals with a paw withdrawal latency onthe ligated side of <13 s in both trials. The animals are then randomlyassigned to groups consisting of 8-10 animals per group The animals aretreated with experimental compounds. The ability of the compounds toextend the latency to paw withdrawal is determined at 1, 3 and/or 5 hpost-treatment.

2. Spinal nerve (L5/L6) Ligation

Adult male Spraque-Dawley rats weighing 150 g-180 g are used in allexperiments. Animals are placed two per cage and allowed free access tofood and water. The cages are housed in rooms that are temperature andhumidity controlled and maintained on a 12-h light/dark cycle.

Spinal nerve ligation is performed on rats anesthetized with sodiumpentobarbital (70 mg/kg, ip). L5 and L6 spinal nerves are ligated with a6.0 silk suture distal to the dorsal root ganglia. The muscle is thenclosed in layers using wound clips and the animals allowed to recoverfor a period of at least 5 days before testing.

Tactile allodynia is evaluated in spinal nerve ligated animals with acalibrated series of eight von Frey filaments as follows. The rats areplaced in clear plastic cages (H: 5′, L: 10″, W: 4⅝″) fitted with a wiremesh floor and allowed to acclimate for 15 min. The following filaments(log 10 of the bending force (g)) are employed to test for allodynia:0.4, 0.7, and 1. 2, 2.0, 3.6, 5.5 8.5 and 15.1 g. Filaments of greaterforce are not used since these alone would physically lift the paw. Eachfilament is applied once to the mid-plantar surface of the affectedhindpaw in a perpendicular fashion aid depressed slowly (4-6 s) untilbending occurred. From the overall pattern of responses a 50% gramwithdrawal threshold is calculated according to the following formula:

50% withdrawal threshold (g)=10(10[^(xf−kd))/10000

where xf is the value (in log units) of the final von Frey hair used: kis the pattern value and d is the mean difference in log units) betweenfilaments: 0.223.

For each experiment, animals are first pre-screened in order to selectfor animals displaying clear signs of tactile allodynia, i.e. animalswith a 50% gram paw threshold of <4-g on the ligated side. The animalsare then randomly assigned to groups consisting of 8-10 animals pergroup. The 50% gram paw withdrawal threshold is then determined 60 minpost-treatment.

Model of Nocioceptive Pain (The “Tail Flick” assay)

The “tail flick” assay is a modification of the method of Ther, L., etal (1963. Zur pharmakodynamischen Wirkiiung der optischen isomeren desmethadons Dtsh Apoth Zig, 103;514). Adult male Spraque-Dawley ratsweighing 150-180 g are used in experiments. Animals are housed two percage and allowed free access to food and water. The cages are housed inrooms that are temperature and humidity controlled and maintained on a12-h light/dark cycle.

Each experimental group consists of 8-10 animals (180 g-220 g). Theanimals are loosely wrapped, individually, in a thin cotton towel withthe head covered and tail exposed. Each animal is placed on a platformwith the tail position in a shallow groove and a focused beam of lightdirected at the tail from above, approximately 2.5-cm from the tip.Movement of the tail from the groove allows the beam of light to hit asensor, formerly covered by the tail, which then automatically switchesoff the beam and stops the timer. The duration of time required for thetail response after exposure to the thermal stimulus is considered thetail response latency time. The maximum time allowed is 10 s in order toprevent tissue damage. Rats are tested once to determine the pre-dosetail response latency following which they are then dosed and againtested for their tail response latency at 60 min post-dose.

II. CENTRAL NERVOUS SYSTEM, (CNS) DISORDERS A. Depression

Antidepressant activity of compounds is tested in rats following themethod of Porsolt, R D., et al (1978, Eur. J. Pharmacol. 47:379. Adultmale Spraque-Dawley rats weighing 150 g-180 g are used in experiments.Animals are placed two per cage and allowed free access to food andwater. The cages are housed in temperature and humidity controlled roomsand maintained on a 12-h light/dark cycle.

The rats (150 g-180 g) are forced to swim in an escape-proof cylinder.After an initial period of vigorous activity they adopt a readilyidentified immobile posture which is used as a model of humandepression. The ability of experimental compounds to increase the periodof time elapsing before animals become immobile is determined.

B. Bipolar Disorder

Current antidepressant therapy, which is based on the inhibition ofbiogenic amine uptake, can be evaluated in vitro. Both rat brainsynaptosomes and washed human platelets are used in these studies. Theinhibition of 5-HT, dopamine, and noradrenaline uptake in these systemsis a measure of the efficacy of the therapeutic compounds.

1. Cells

The effect of test compounds on 5-HT transport in washed human plateletsis evaluated following the method of Southam, E., et al (1998, Eur. J.Pharmacol. 358:19). Human blood is obtained from volunteers andplatelets isolated by centrifugation, washed, and resuspended (3×10⁵/ul)in cold (4° C.) HEPES buffer (pH 7.4) consisting of 5.0 mM HEPEScontaining 140 mM NaCl, 2.82 mM KCL, 0.74 mM KH₂PO₄, 5.5 mM NaHCO₃, 1 mMCaCl₂, 0.5 mM MgSO₄ and 5.1 mM glucose. 10 uM pargyline is added toinhibit monamine oxidase activity.

Cells are obtained from adult male Lister hooded rat cortex (5-HT andnoradrenaline uptake) or striatum (dopamine uptake). The cortex andstriatum are homogenized in 0.32 M sucrose solution and syntaptosomesisolated by centrifugation before being gently suspended in cold (4₀C)pre-gassed (5% CO₂, 95% O₂) Krebs solution containing 115 mM NaCl, 4.97KCL, 1.2 mM KH₂PO₄, 5.5 mM NaHCO₃, 1.0 mM CaCl₂, 1.22 mM MgSO₄, 11.1 mMglucose, 0,01 mM pargyline. The protein concentration (0.2-0.5 mg/ml ofthe crude synaptosomal preparation, is determined following the methodof Bradford (Bradford, M M., 1976. Anal. Biochem. 72:248).

Biogenic Amine Uptake.

190 ul of either the platelet or synaptosome preparation are added tosolutions containing 800 ul of one of three tritiated biogenicamines: 1) [³H] 5-HT (final concentration 20 mM): or 2) [³H]noradrenaline (50 nM); or 3) [³H] dopamine (20 nM). Test compounds areadded and the mixtures incubated for 10 min at 37° C. The mixture ofcells, biogenic amines, and test substances are individually filteredthrough pre-wetted Whatman GF/B filter paper. Then the filter paper waswashed 3 times with ice-cold buffer to stop the uptake of the tritiatedamines. Liquid scintillation counting assesses the radioactivitycaptured on the filter paper. Non-specific uptake is determined andsubsequently subtracted from counts. Data points represent the mean±SEMof at least four different assays. Each assay point is performed intriplicate and expressed as a percentage of controls (also performed intriplicate). IC₅₀s are generated by calculating the geometric mean(number (n) and 95% confidence interval (CI₉₅) indicated in parentheses)of values estimated by fitting a sigmoidal model of the following formusing a non-linear curve fit based on the algorithm of Marquard (1963,J. Soc. Indust. Appl. Math. 11:431):$Y = \frac{\left( {a - d} \right)}{1 + {\left( {x/c} \right)b}}$

where, y=raw counts: x=concentration of compound: a=lower asymptote:d=upper asymptote, b=Hill slope and c=IC₅₀. The assumption is thatuptake will be depressed to non-specific levels at infinitely highconcentrations.

Neurodegeneration

The neuroprotective effect of the compounds is tested in vitro in amodel of neurodegeneration. In this model cytotoxicity is induced byglutamate as described by Huettner, J E and Baughman, R W. (1986. J.Neurosci. 6:3044). Briefly, rat pups aging from newborn to 1 dayweighing from 6 g to 8 g are anesthetized with chloral hydrate. Thecortices with hippocami attached are removed and placed in Cl freedissociation medium supplemented with 1-mM kynurenic acid and 10 mMMgSO₄. The tissue is cleared of meninges, washed, and incubated for 20min at 37° C. in dissociation medium containing 10 units/ml papain(Wortington), a digestive enzyme. The tissue is then incubated for three5-min periods at 37° C. in isotonic. medium containing 10-mg/ml trypsininhibitor to stop the reaction.

The cells are dissociated by trituration and resuspended in growthmedium (GM) consisting of Eagles minimum essential medium (MEM)supplemented with 5% fetal bovine serum, 5% defined supplemented calfserum (hyclone), 50 mM glucose, 50 U/ml penicillin/streptomycin andserum extender (Collaborative Research). The cells (5×10⁵/ml) arealiquoted (0.1 ml/well) into the wells of 96 well plates pre-coated withpoly-D-ysine (0.5 mg/ml) and laminin (2 ug/nl) (Collaborative Research)so that a final density of 5×10⁴ cells per well 1s achieved. The cellsare maintained at 37° C. in a humidified incubator in an atmosphere of5% CO₂ in air.

Fresh media is added to the cultures by removing one half of the mediaand adding the equivalent volume of new media twice weekly for 15-16days.

Experiments are performed on cultures in which the neurons are at auniform density. Cultures are washed three times in a modification ofthe medium used by Choi et al (1987, J. Neurosci. 7:257) consisting ofHEPES-buffered control salt solution (CSS) containing 10 mM HEPESbuffered at pH 7.4. In all experiments the cells are incubated with aneurotoxic concentration of glutamate (500 uM). Test compounds, dilutedin CSS are added to the cultures in 2-fold serial dilutions. Controlcultures are incubated with CSS alone, or serial dilutions of testcompounds in CSS alone, or 500 uM glutamate. The wells are washed threetimes with CSS and 100 ul aliquots of glucose enriched MED are added toall wells. The plates are maintained overnight at 37° C. in anatmosphere of 5% CO₂ in air.

Glutamate-induced death of neurons is measured by determining the levelsof lactate dehydrogenase (LDH) released into the medium by dead anddying neurons 2448 hours following glutamate insult (Koh and Choi, 1987,J. Neurosci. Methods, 20:83). Media samples are collected from all wellsand assayed for LDH according to the protocol suggested by MolecularDevices Applications Bulletin, 012-A using the Molecular Devices KineticMicroplate Reader. Results are normalized to the LDH values obtained inthe. glutamate alone controls.

The concentration of test compound resulting in a 50% inhibition ofrelease of LDH is the ED₅₀.

Cerebrovascular Ischemia

Following the method of O=Neill, M J et al. (1997, Eur. J. Pharmacol.332:121) male Mongolian gerbils, at least 3 months old and weighing inexcess of 60 g, are used in these in vivo experiments. The animals aremaintained in standard lighting conditions and food and water are.available ad libitum. The animals are anesthetized with 5%halothane/oxygen mixture and maintained using 2% halothane deliveredwith oxygen at 1 L/min via a face mask throughout the operation. Througha midline cervical incision, both common carotid arteries are exposedand freed from surrounding connective tissue. In animals to be renderedischemic both common carotid arteries are clamped for 5 min. to occludethe blood flow. At the end of the occlusion period blood flow wasre-established. In sham operated animals the arteries are exposed butnot occluded. The wound is then sutured and the animals allowed torecover. Throughout the surgery body temperature is maintained at 37° C.using a “K-TEMP” temperature controller/heating pad (InternationalMarket Supply) and brain temperature are maintained using a heatinglamp. After surgery the animals are placed in a four compartmentthermacage (Beta Medical and Scientific) which maintained theenvironmental temperature at 28° C. and rectal temperatures are measuredfor a 6 h period after occlusion.

The doses of compounds were selected based on previous work andadministered a various times prior to during and after the occlusion. 5days after surgery the animals are perfused transcardially with 30 ml of0.9% saline followed by 100 ml of 10% buffered formalin solution. Thebrains are removed and placed in 10% formalin for 3 days processed andembedded in paraffin wax. 5 um coronal sections are taken 1.5-1.9 mmcaudal to the bregma in the anterior hippocampus using a sledgemicrotome(Leitz 1400). The slices were stained with hemoatoxylin and eosin andthe neuronal density in the CA1 subfield of the hippocampus was measuredusing a microscope with grid lines (0.05 mm×0.05 mm). The neuronaldensity is expressed as neuronal density per mm CA1 hippocampus.

Epilepsy: Bioassays of the Effect of Compounds on Experimentally InducedSeizures and Convulsions.

Following the method of Dalby, N., et al (1997, Epilepsy Research28:63). Groups of animals (10 animals/group) receive ip injections ofeither vehicle or test compounds at a variety of concentrations prior toinducing seizures or convulsions as described below.

1. Seizures Induced by Maximal Electroshock

Male NMRI mice (20±2 g) of either sex are maintained in groups of 40.The cages (59×38×20 cm) are placed in a room at 22° C. with a relativehumidity of 55% in a 12 h/12 h normal light/dark cycle with ad libitumaccess to food and water. The mice are stimulated by corneal electrodesfrom a Hugo Sachs stimulator (type 207) with 50 mA, 60 Hz AC, for 0.2 s.The animals are observed for tonic hindlimb extension following 10 safter stimulation. An ED₅₀ value is determined as the dose of ligandprotecting 50% of the animals against tonic hindlimb extension.

2. Seizures Induced by Sound.

DBA/2 mice of either sex (8+1 g) 18-21 days old are individually exposedto a 111 db sinusoidal tone at 14 kHz for 30 s and observed for thepresence of clonic and tonic convulsions during this period. An ED₅₀value is determined as the dose of ligand (umol/kg) protecting 50% ofthe animals from clonic or/and tonic convulsions.

3. Pentylentetrazol (PTZ) Induced Convulsions

Male NMRI mice (20±2 g) are injected subcutaneously with 160 mg/kg ofPTZ to induce tonic convulsions. The mice are observed for the following15 min and time to tonic convulsions is noted for each animal. For PTZinduced clonic convulsions, a dose of PTZ (120 mg/kg) is administeredsubcutaneously, and the animals observed for the following 30 min andtime to clonic convulsions is noted for each animal. ED₅₀ values aredetermined as the dose of ligand which protects 50% of the animalsagainst clonic or tonic convulsion.

4. 6.7-dimethoxy-4ethyl-β-carboline-3-carboxylate (DMCM) InducedConvulsions.

Male NMRI mice (20±2 g) receive DMCM (18 mg/kg) and are observed for 15min following injection for the presence of clonic and tonic convulsionsand death. An ED₅₀ value is determined as the dose of ligand protecting50% of the animals against clonic or/and toxic convulsions.

Schizophrenia

Subjects diagnosed with schizophrenia are selected from a group ofinpatients. All subjects give written informed consents to participate.

After a 2 week baseline assessment period, subjects are randomlyassigned to receive, under double-blind conditions, either experimentalcompounds (dissolved in water) or placebo (glucose in water). Eachpatient undergoes a 2-week adjunctive treatment washout period afterwhich he/she crossed over to the alternative substance for a further 6weeks. Experimental compounds are administered at a variety ofconcentrations. The only other medications allowed during the study wastrilexyphenidyl (2-5 mg/day) for treatment of extrapyramidal symptomsand chloral hydrate (250-750 mg/day on PRN basis) for treatment ofinsomnia or agitation. For patients needing antiparkinsonian medication,trihexyphenidyl dose was kept constant throughout the study.

Symptoms and extrapyramidal side effects are assessed starting fromweek-2, biweekly throughout the study, using Positive and NegativeSymptom Scale (PANSS), the Simpson-Angus Scale for ExtrapyramidalSymptoms (SAS) and Abnormal Involuntary Movement Scale (AIMS). Patientsrequiring, at any point during the study, neuroleptic dose increases arewithdrawn and appropriate treatment instituted. Withdrawl decisions arebased on clinical evaluations and coincide with an increase of at least30% on the PANSS score.

Physical complaints and status are monitored daily. Hematology, bloodchemistry, liver and kidney function, laboratory measures are assessedbiweekly. Blood samples for the assessment of serum levels ofexperimental compounds are obtained at baseline and at the end of studyweeks 6 and 14. Blood drawings are performed before breakfast and firstdaily administration of medication. Serum compound levels weredetermined by HPLC.

The anti-anxiety effects of compounds are studied in vivo following themethod of Costall, B., et al (1987, Neuropharmacol. 26:195). Briefly,mice that tend to explore a novel environment, are placed I n atwo-chambered system in which they can freely move between a brightlylit open field and a dark corner. The animals are averse to moving intothe bright area. The ability of compounds to suppress anxiety aboutmoving into the bright light is determined.

Briefly, naïve male albino mice with a weight between 18 g and 25 g areplaced into a testing apparatus consisting of a light and dark chamberdivided by a photocell-equipped zone. A polypropylene animal cage,44×21×21 cm is darkened with black spray over one-third of its surface.A partition containing a 13-cm long×5-cm high opening separates the darkone third from the bright two thirds of the cage. The cage rests on anAnimex activity monitor which counts total locomoter activity. Anelectronic system using four sets of photocells across the partitionautomatically count movements through the partition and clocks the timespent in the light and dark compartments. The animals are treated 30 minbefore the experiment with the test drugs or the vehicle intraperitoneal and are then observed for 10 min.

Dose-response curves are obtained and the number of crossings throughoutthe partition between the light and dark chamber are compared with totalactivity counts during the 10 minutes.

Migraine: Plasma Extravasation Model

Plasma extravasation is a sequela of neurogenic inflammation within thedura mater and the mechanism involved in the production of migraneheadache. Plasma extravasation may be experimentally produced in thedura by electrical stimulation of the trigeminal ganglion. Theprotective effect of therapeutic compounds on extravasation can beevaluated in such models.

Thus, following the method of Shepheard, S L, et al., (1995,Neuropharmacology, 34(3):255) male Sprague-Dawley rats (180 g-230 g) areanaesthetized with pentobarbitone sodium (60 mg kg⁻1, ip) and anaethesiamaintained by giving supplementary doses (10 mg kg⁻¹, iv) as required. Afemoral vein and artery are cannulated for iv injections and monitoringof arterial blood pressure respectively. Rats are placed in a sterotaxicframe and burr holes drilled 3.2 mm posterior and 2.8 mm lateral frombregma to allow placement of bipolar concentric stimulating electrodes(NE 200X, Clark Electromedical). Test compounds dissolved in H₂O or H₂Oalone are administered (n=8-10 per group) at time t=0 min. Theradioactive plasma marker, ¹²⁵I bovine serum albumen (70 uCi kg⁻1) wasadministered at t=5 min. The electrodes are lowered 9.5 mm below thesurface of the dura and at t=10 min the right trigeminal ganglion isstimulated with 0.6 mA constant current, 5 mSec duration square wavepulse, 5 Hz for 5 min. At t=20 min the animals are killed byexsanguination and the dura dissected from both sides of the skull. Thearea of dura immediately surrounding the sites of electrode penetrationis discarded. Correct electrode placement is confirmed by the presenceof electrode marks in the trigeminal ganglion. Samples of extracranialtissues (conjunctiva, eyelid and lip) innervated by the trigeminal nerveare also taken. Tissues are rinsed, dried overnight weighed and thencounted for radioactivity.

The counts mg⁻¹ dry weight of tissue samples from the stimulated andunstimulated sides are calculated and results expressed as anextravasation ratio, ie the ratio of the stimulated to unstimulatedsides. For calculation of an ID₅₀ value, (dose producing 50%extravasation) an inhibition curve is fitted to the data using “Grafit”curve-fitting software (Erithacus software, UK).

STATISTICS

Pain Models

All group comparison data are analyzed using a Kruskal-Wallis one wayanalysis of variance (ANOVA) followed by a pairwise comparison betweenvehicle and each drug-treated group with a Dunnett=s t-test on theranked data Ligand effects are considered to be statisticallysignificant only if they are different from both the pre-dose data andthe vehicle data (at that time point) at the P<0.05 level.

Seizure and Convulsion Models

ED₅₀ values and 95% confidence intervals (CI_(95%)) in all tests areobtained using log-probit method.

In Vivo Model of Ischemia

Statistical analysis of histological data is carried out using ANOVAfollowed by Student=s t-test with Bonferoni corrections using P<0.05 asthe level of significance.

Schizophrenia

To assess treatment responses to compounds, rmANOVA. In order to assessthe possibility that treatment order affected overall results, rmNAOVAof negative symptoms by treatment phase and week are covaried fortreatment order.

Pharmaceutical Formulations

When employed as pharmaceuticals, the compounds of formula I are usuallyadministered in the form of pharmaceutical compositions. This inventiontherefore provides pharmaceutical compositions which contain, as theactive ingredient, one or more of the compounds of Formula I above or apharmaceutically acceptable salt thereof and one or morepharmaceutically acceptable excipients, carriers, diluents, permeationenhancers, solubilizers and adjuvants. The compounds may be administeredalone or in combination with other therapeutic agents (e.g., otherantihypertensive drugs, diuretics and the like). Such compositions areprepared in a manner well known in the pharmaceutical art (see, e.g.,Remington's Pharm. Sci., Mack Publishing Co., Philadelphia, Pa., 17^(th)Ed. (1985) and “Modem Pharm.,” Marcel Dekker, Inc., 3^(rd) Ed. (G. S.Banker & C. T. Rhodes, Eds.).

The compounds of Formula I may be administered by any of the acceptedmodes of administration of agents having similar utilities, for example,by oral, parenteral, rectal, buccal, intranasal or transdermal routes.The most suitable route will depend on the nature and severity of thecondition being treated. Oral administration is a preferred route forthe compounds of this invention. In making the compositions of thisinvention; the active ingredient is usually diluted by an excipient orenclosed within such a carrier which can be in the form of a capsule,sachet, paper or other container. When the excipient serves as adiluent, it can be a solid, semi-solid, or liquid material, which actsas a vehicle, carrier or medium for the active ingredient. Thus, thecompositions can be in the form of tablets, pills, powders, lozenges,sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups,aerosols (as a solid or in a liquid medium), ointments containing, forexample, up to, 10% by weight of the active compound, soft and hardgelatin capsules, suppositories, sterile injectable solutions, andsterile packaged powders. Pharmaceutically acceptable salts of theactive agents may be prepared using standard procedures known to thoseskilled in the art of synthetic organic chemistry and described, e.g.,by J. March, Advanced Organic Chem. Reactions, Mechanisms and Structure,4^(th) Ed. (N.Y.: Wiley-Interscience, 1992).

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents.

The compositions of the invention can be formulated so as to providequick, sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.Controlled release drug delivery systems for oral administration includeosmotic pump systems and dissolutional systems containing polymer-coatedreservoirs or drug-polymer matrix formulations. Examples of controlledrelease systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525;4,902,514; and 5,616,345. Another preferred formulation for use in themethods of the present invention employs transdermal delivery devices(“patches”). Such transdermal patches may be used to provide continuousor discontinuous infusion of the compounds of the present invention incontrolled amounts. The construction and use of transdermal patches forthe delivery of pharmaceutical agents is well known in the art. See,e.g., U.S. Pat. Nos. 5,023,252; 4,992,445 and 5,001,139. Such patchesmay be constructed for continuous, pulsatile, or on demand delivery ofpharmaceutical agents.

The compositions are preferably formulated in a unit dosage form. Theterm “unit dosage forms” refers to physically discrete units suitable asunitary dosages for human subjects and other mammals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect, in association with a suitablepharmaceutical excipient (e.g., a tablet, capsule, ampoule). The activecompound is effective over a wide dosage range and is generallyadministered in a pharmaceutically effective amount. Preferably, fororal administration, each dosage unit contains from 1-250 mg of acompound of Formula I, and for parenteral administration, preferablyfrom 0.1 to 60 mg of a compound of Formula I or a pharmaceuticallyacceptable salt thereof. It will be understood, however, that the amountof the compound actually administered will be determined by a physician,in the light of the relevant circumstances, including the condition tobe treated, the chosen route of administration, the actual compoundadministered and its relative activity, the age, weight, and response ofthe individual patient, the severity of the patient's symptoms, and thelike.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules.

The tablets or pills of the present invention may be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as corn oil,cottonseed oil, sesame oil, coconut oil, or peanut oil, as well aselixirs and similar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be inhaled directly from thenebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices which deliver the formulationin an appropriate manner.

The following formulation examples illustrate representativepharmaceutical compositions of the present invention.

FORMULATION EXAMPLE 1

Hard gelatin capsules containing the following ingredients are prepared:

Quantity Ingredient (mg/capsule) Active Ingredient 30.0 Starch 305.0Magnesium stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules in340 mg quantities.

FORMULATION EXAMPLE 2

A tablet formula is prepared using the ingredients below:

Quantity Ingredient (mg/tablet) Active Ingredient 25.0 Cellulose,microcrystalline 200.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0

The components are blended and compressed to form tablets, each weighing240 mg.

FORMULATION EXAMPLE 3

A dry powder inhaler formulation is prepared containing the followingcomponents:

Ingredient Weight % Active Ingredient 5 Lactose 95

The active ingredient is mixed with the lactose and the mixture is addedto a dry powder inhaling appliance.

FORMULATION EXAMPLE 4

Tablets, each containing 30 mg of active ingredient, are prepared asfollows:

Quantity Ingredient (mg/tablet) Active Ingredient 30.0 Starch 45.0Microcrystalline cellulose 35.0 Polyvinylpyrrolidone 4.0 (as 10%solution in sterile water) Sodium carboxymethyl starch 4.5 Magnesiumstearate 0.5 Talc 1.0 Total 120.0

The active ingredient, starch and cellulose are passed through a No. 20mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders, which are thenpassed through a 16 mesh U.S. sieve. The granules so produced are driedat 50° C. to 60° C. and passed through a 16 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 30 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 120 mg.

FORMULATION EXAMPLE 5

Capsules, each containing 40 mg of medicament are made as follows:

Quantity Ingredient (mg/capsule) Active Ingredient 40.0 Starch 109.0Magnesium stearate 1.0 Total 150.0

The active ingredient, starch, and magnesium stearate are blended,passed through a No. 20 mesh U.S. sieve, and filled into hard gelatincapsules in 150 mg quantities.

FORMULATION EXAMPLE 6

Suppositories, each containing 25 mg of active ingredient are made asfollows:

Ingredient Amount Active Ingredient 25.0 mg Saturated fatty acidglycerides to 2,000.0 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

FORMULATION EXAMPLE 7

Suspensions, each containing 50 mg of medicament per 5.0 mL dose aremade as follows:

Ingredient Amount Active Ingredient 50.0 mg Xanthan gum 4.0 mg Sodiumcarboxymethyl cellulose (11%) Microcrystalline cellulose (89%) 50.0 mgSucrose 1.75 g Sodium benzoate 10.0 mg Flavor and Color q.v. Purifiedwater to 5.0 ml

The active ingredient, sucrose and xanthan gum are blended, passedthrough a No. 10 mesh U.S. sieve, and then mixed with a previously madesolution of the microcrystalline cellulose and sodium carboxymethylcellulose in water. The sodium benzoate, flavor, and color are dilutedwith some of the water and added with stirring. Sufficient water is thenadded to produce the required volume.

FORMULATION EXAMPLE 8

Quantity (mg/ Ingredient capsule) Active Ingredient 15.0 mg Starch 407.0mg Magnesium stearate 3.0 mg Total 425.0 mg

The active ingredient, starch, and magnesium stearate are blended,passed through a No. 20 mesh U.S. sieve, and filled into hard gelatincapsules in 425.0 mg quantities.

FORMULATION EXAMPLE 9

A subcutaneous formulation may be prepared as follows:

Ingredient Quantity Active Ingredient 5.0 mg Corn Oil 1.0 mL

Frequently, it will be desirable or necessary to introduce thepharmaceutical composition to the brain, either directly or indirectly.Direct techniques usually involve placement of a drug delivery catheterinto the host's ventricular system to bypass the blood-brain barrier.One such implantable delivery system used for the transport ofbiological factors to specific anatomical regions of the body isdescribed in U.S. Pat. No. 5,011,472 which is herein incorporated byreference.

Indirect techniques, which are generally preferred, usually involveformulating the compositions to provide for drug latentiation by theconversion of hydrophilic drugs into lipid-soluble drugs. Latentiationis generally achieved through blocking of the hydroxy, carbonyl,sulfate, and primary amine groups present on the drug to render the drugmore lipid soluble and amenable to transportation across the blood-brainbarrier. Alternatively, the delivery of hydrophilic drugs may beenhanced by intra-arterial infusion of hypertonic solutions which cantransiently open the blood-brain barrier.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

All of the publications, patent applications and patents cited in thisapplication are herein incorporated by reference in their entirety tothe same extent as if each individual publication, patent application orpatent was specifically and individually indicated to be incorporated byreference in its entirety.

What is claimed is:
 1. A compound of the formula:

and pharmaceutically acceptable salts thereof; wherein each R²⁰ isindependently hydrogen, methyl or ethyl; and X is a linker of theformula: —X′—Z—(Y′—Z)_(m)—Y″—Z—X′—  in which: m is an integer of from 0to 20; X′ at each separate occurrence is selected from the groupconsisting of —O—, —S—, —NR—, —C(O)—, —C(O)O—, —C(O)NR—, —C(S), —C(S)O—,—C(S)NR— or a covalent bond where R is as defined below; Z at eachseparate occurrence is selected from the group consisting of alkylene,substituted alkylene, cycloalkylene, substituted cylcoalkylene,alkenylene, substituted alkenylene, alkynylene, substituted alkynylene,cycloalkenylene, substituted cycloalkenylene, arylene, heteroarylene,heterocyclene, or a covalent bond; Y′ and Y″ at each separate occurrenceare selected from the group consisting of:

 —S—S— and a covalent bond;  in which: n is 0, 1 or 2; and R, R′ and R″at each separate occurrence are selected from the group consisting ofhydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl,alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic.
 2. Thecompound of claim 1, wherein each R²⁰ is hydrogen.
 3. The compound ofclaim 1, wherein each R²⁰ is methyl.
 4. The compound of claim 1, whereineach R²⁰ is ethyl.
 5. A compound of the formula:

and pharmaceutically acceptable salts thereof; wherein each R²⁰ isindependently hydrogen, methyl or ethyl; and X¹ is a linker of theformula:

 wherein each R^(a) is independently selected from the group consistingof a covalent bond, alkylene, substituted alkylene and arylene; eachR^(b) is independently selected from the group consisting of hydrogen,alkyl and substituted alkyl; and n′ is an integer ranging from 1 toabout
 20. 6. The compound of claim 5, wherein each R²⁰ is hydrogen. 7.The compound of claim 5, wherein each R²⁰ is methyl.
 8. The compound ofclaim 5, wherein each R²⁰ is ethyl.
 9. A compound of the formula:

and pharmaceutically acceptable salts thereof; wherein said compound isselected from the group consisting of compounds wherein X² and R²⁰ areas follows: X² R²⁰ —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— H; —(CH₂)₃—O—(CH₂)₃— H;—CH₂—Z—CH₂— where Z = 1,3-cyclohexyl H; —CH₂—CH(OH)—CH₂— H;

H; —CH₂—Z—CH₂— where Z = 1,4-cyclohexyl H; —(CH₂)₅— H; —(CH₂)₆— H;—(CH₂)₂—S—(CH₂)₂— H; —(CH₂)₃—O—(CH₂)₁₀—O—(CH₂)₃— H; —(CH₂)₃—Z—(CH₂)₃—where Z = H; 2,4,8,10-tetraoxa-spiro[5.5]undecan-3,9-yl—(CH₂)₃—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₃— H; —(CH₂)₃—O—(CH₂)₄—O—(CH₂)₃— H;—(CH₂)₃—N(CH₃)—(CH₂)₃— H; —CH₂—Z—CH₂— where Z = trans-1,4-cyclohexyl H;—CH₂—CH(CH₃)—CH₂—C(CH₃)₂—(CH₂)₂— H; —Z— where Z = 2,7-[9H-fluorene] H;—Z—C(CH₃)₂—Z—C(CH₃)₂—Z— where Z = 1,4-phenyl H; —Z—CH₂—Z— where Z =1,4-phenyl H; —(CH₂)₂—N(CH₃)—(CH₂)₂— H; —(CH₂)₉— Me; —CH₂—Z—CH₂— where Z= 1,4-phenyl Me; —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Me; —CH₂—CH═CH—CH₂— (transisomer) Me; —CH₂—Z—CH₂— where Z = (-)trans-2,2-dimethyl- Me;[1,3]dioxolan-4,5-yl —(CH₂)₁₀— Me; —(CH₂)₁₁— Me; —(CH₂)₁₂— Me; —(CH₂)₁₆—Me; —CH₂—CH(OH)—CH₂— Me; —CH₂—C═C—CH₂— Me; —(CH₂)₇— Me —(CH₂)₈— Me—CH₂—Z—CH₂— where Z = 2,3-quinoxalinyl Me —CH₂—CH(CH₂OH)— Me—CH₂—C(CH₂)—CH₂— Me —Z— where Z = 4,5-[1,3]dioxolan-2-one Me —CH₂—Z—CH₂—where Z = 1,3-phenyl Me —(CH₂)₂—O—CH₂—O—(CH₂)₂— Me —(CH₂)₄—O—(CH₂)₄— Me—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Me —(CH₂)₂—O—C(O)—O—(CH₂)₂— Me—CH₂—CH[C(O)—O—CH₂CH₃]— Me —CH₂—C(O)—O—(CH₂)₂—O—C(O)—CH₂— Me—(CH₂)₂—NH—C(O)—C(O)—NH—(CH₂)₂— Me —(CH₂)₃— Me—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Et; —CH₂—Z—CH₂— where Z = (-)trans-2,2- Et;dimethyl-[1,3]dioxolan-4,5-yl —(CH₂)₁₀— Et; —(CH₂)₁₁— Et; —(CH₂)₁₂— Et;—(CH₂)₁₆— Et; —(CH₂)₇— Et; —CH₂—Z—CH₂— where Z = 1,3-phenyl Et;—CH₂—Z—CH₂— where Z = 1,4-phenyl Et; —(CH₂)₂—O—CH₂—O—(CH₂)₂— Et;—(CH₂)₄—O—(CH₂)₄— Et; —(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂— Et;—CH₂—C(O)—O—(CH₂)₂—O—C(O)—CH₂— Et; —(CH₂)₃— Et; —CH₂—Z—CH₂— where Z =2,5-thiophenyl H; —CH₂—Z—O—(CH₂)₂—O—Z—CH₂— where Z = 1,2-phenyl H;—CH₂—Z—O—(CH₂)₆—O—Z—CH₂— where Z = 1,2-phenyl H;—CH₂—Z—O—(CH₂)₃—O—Z—CH₂— where Z = 1,2-phenyl H; —CH₂—Z—CH₂— where Z =2,3-thiophenyl H; —CH₂—Z—CH₂— where Z = 2,6-pyridinyl H; —CH₂—Z—CH₂—where Z = 2-hydroxy- H; 5-methyl-phenyl-1,3-yl —CH₂—Z—O—(CH₂)₂—O—Z—CH₂—where Z = H; 4-methoxy-phenyl-1,3-yl —CH₂—Z—CH₂— where Z =4-hydroxy-phenyl-1,3-yl H; —CH₂—Z—CH₂— where Z = 2,2′-dihydroxy-3,3′- H;dimethoxy-biphenyl-5,5′-yl —CH₂—Z—O—Z—CH₂— where Z = 1,2-phenyl H; and—CH₂—Z—CH₂— where Z = 1,3-phenyl H.


10. The compound of claim 9, wherein each R²⁰ is hydrogen, and X² is—CH₂—Z—CH₂— where Z is pyrridin-2,6-diyl.
 11. A pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient and atherapeutically effective amount of a compound of any of claims 1-10, ora pharmaceutically acceptable salt thereof.
 12. A method of treatingpain in a mammal, the method comprising administering to said mammal aneffective amount of a pharmaceutical composition according to claim 11.13. A method of modulating the activity of a sodium channel in abiological tissue, the method comprising contacting a biological tissuehaving a sodium channel with a sodium channel modulating amount of acompound of any of claims 1-10, or a pharmaceuticall acceptable saltthereof.